• Title/Summary/Keyword: Microcosm soil test

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Characterization of Diesel Oil-Degrading Bacteria (디젤유 분해균주의 특성 및 토양배양)

  • 안민정;한윤전;임현섭;최기현;권오범;정병철
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.108-113
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    • 2003
  • Diesel oil-degrading bacterial strains were isolated from diesel oil contaminated soil and called HS series (HS1, HS2 and HS3). These strains were identified as Acinetobacter sp. (HS1) and Pseudomonas sp. (HS2 and HS3) based on Biolog test, cellular fatty acid composition, and 16S rDNA sequence analysis. These strains were coltivated in liquid minimal media containing 2% diesel oil, and diesel oil-degrading activity was measured. As result, all strains degraded over 70% of total diesel oil. But PAH (polycyclic aromatic hydrocarbon)- and pris- tane-degrading rate of these strain was below 20% of total PAH and pristane. The HS 1 strain showed highest hydrophobicity and low emulsifying activity among the experimental strains and high diesel oil-degrading activity. From the above-mentioned result, microcosm experiment was performed with the HS1 strain. The HS1 strain showed a degrading activity of over 80% of total diesel oil in microcosm test. And microbial activity was correlated to diesel oil-degrading activity. Therefore, it is suggested that the HS1 strains could be effectively used for the bioremediation for diesel oil.

Asswssment of natural attenuation for Gasoline contaminated soil under various reducing conditions

  • 오인석;이시진;장순웅
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.255-259
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    • 2004
  • 본 연구에서는 유류오염 토양현장샘플을 이용한 실험실 규모의 회분식 실험을 통해 다양한 환원조건하에서 BTEX와 MTBE의 자연정화 평가 및 현장 생복원 기술 적용 타당성을 검토하는데 있다. 첫째, 오염토양과 비오염 토양에서 충분한 전자수용체의 존재는 유류오염물질의 생분해능을 증가시킬 수 있었음 알았다. 둘째, 같은 토양이라도 전자수용체의 종류와 유류오염물질의 특성에 따른 생분해능의 다양성을 확인할 수 있었다. 셋째, BTEX에 비해 MTBE의 분해가 매우 느림을 확인하였다. 본 실험은 아직 초기 단계의 실험으로 현장 조건을 충분히 만족 시키지는 못한다. 추가적인 pH 변화에 따른 전자수용체의 이용 특성 및 토양-지하수 microcosm test, 혐기성 생분해 부산물인 $CO_2$, $N_2$ 또는 $N_2$O, TBA의 연구가 실시되어야 하겠다.

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A Study Bioremediation of Tidal Flat by Microorganism in Pilot Scale Test (환경정화 미생물에 의한 갯벌의 생물학적 정화에 대한 파일럿 규모의 연구)

  • Choi, Hye Jin;Han, Young Sun;Park, Doo Hyun;Oh, Bo Young;Hur, Myung Je;Jo, Nam-Gyu;Kim, Young Hee;Kim, Jong-Guk
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1110-1117
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    • 2014
  • Tidal flats are continuously contaminated by human activities. This study assessed the bioremediation efficiency of tidal flat soil using microcosm reactors and microorganisms originating from the tidal area. We screened 135 bacterial strains that produce extracellular enzymes from the tidal area located in the North port of Incheon bay. Two bacterial strains (Pseudoalteromonas sp. and IC35 Halothiobacillus neapolitanus IC_S22) were selected and used in the microcosm reactors, which were specially designed to functionally mimic the ecological conditions of the tidal flats. Pseudoalteromonas sp. IC35 was selected based on its relatively high activity of the enzymes amylase, cellulose, lipase, and protease. Halothiobacillus neapolitanus IC_S22 was selected for oxidation of sulfur. The M1 and M2 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M2 was first inoculated with Pseudoalteromonas sp. IC35 before the seawater feeding. The initial COD in both the M1 and M2 microcosm reactors was 320 mg/l. The final COD was 21 mg/l (M1) and 7 mg/l (M2). The M3 and M4 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M4 was first inoculated with H. neapolitanus IC_S22. The initial sulfate concentration in both the M3 and M4 microcosm reactors was 660 mg/l, and the maximum sulfate concentration was 1,360 mg/l (M3) and 1,600 mg/l (M4).

Effects of Size of Metal Particles on Soil Microbial Community and Buck Wheat (금속 입자 크기가 토양 미생물 군집과 메밀에 미치는 영향)

  • Kim, Sung-Hyun;Kim, Jung-Eun;Gwak, Young-Ji;Kim, Yun-Ji;Lee, In-Sook
    • Journal of Environmental Science International
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    • v.20 no.4
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    • pp.457-463
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    • 2011
  • This study was carried out to compare the toxicity of nano and micrometer particles with Cu and Zn on soil microbial community and metal uptake of buck wheat. In microcosm system, soil was incubated for 14 days after soil aliquots were artificially contaminated with 1,000 mg/kg Cu, Zn nano and micro particles, respectively. After then, buck wheat was planted in incubating soils and non incubating soils. After 14 days, we compared bioaccumulation of metal, and microbial carbon substrate utilization patterns between incubating soils and non-incubating soils. The enrichment factor (EF) values of incubating samples were greater than non-incubating soils. Dehydrogenase activity had been inhibited by Cu and Zn nanoparticles in non-incubating soil, as well as it had been inhibited by Zn micro particles in incubating soils. Results of biolog test, it was not significant different between nano particles and micro particles. It cannot be generalized that nanoparticles of metal are always more toxic to soil microbial activity and diversity than micrometer-sized particles and the toxicity needs to be assessed on a case-by-case basis.

Monitoring 4-Chlorobiphenyl-Degrading Bacteria in Soil Microcosms by Competitive Quantitative PCR

  • Lee, Soo-Youn;Song, Min-Sup;You, Kyung-Man;Kim, Bae-Hoon;Bang, Seong-Ho;Lee, In-Soo;Kim, Chi-Kyung;Park, Yong-Keun
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.274-281
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    • 2002
  • The competitive quantitative PCR method targeting pcbC gene was developed for monitoring 4-chlorobiphenyl(4CB)-degrading bacteria, Pseudomonas sp. strain DJ-12, in soil microcosms. The method involves extraction of DNA from soil contaminated with 4CB, PCR amplification of a pcbC gene fragment from the introduced strain with a set of strain-specific primers, and quantification of the elec-trophoresed PCR product by densitometry. To test the adequacy of the method, Pseudomonas sp. strain DJ-12 was introduced into both contaminated and non-contaminated soil microcosms amended with 4CB. Pseudomonas sp. strain DJ-12 was monitored and quantified by a competitive quantitative PCR in comparison with 4CB degradation and the result was compared to those obtained by using the conventional cultivation method. We successfully detected and monitored 4CB-degrading bacteria in each microcosm and found a significant linear relationship between the number of 4CB-degrading bacteria and the capacity for 4CB biodegradation. The results of DNA spiking and cell-spreading experiments suggest that this competitive quantitative PCR method targeting the pcbC gene for monitoring 4CB- degrading bacteria appears to be rapid, sensitive and more suitable than the microbiological approach in estimating the capacity of 4CB biodegradation in environmental samples.

Evaluation of Microbial PCE Reductive Dechlorination Activity and Microbial Community Structure using PCE-Contaminated Groundwater in Korea (사염화에틸렌(PCE)으로 오염된 국내 4개 지역 지하수 내 생물학적 PCE 탈염소화 활성 및 미생물 군집의 비교)

  • Kim Young;Kim Jin-Wook;Ha Chul-Yoon;Kwon Soo-Yeol;Kim Jung-Kwan;Lee Han-Woong;Ha Joon-Soo;Park Hoo-Won;Ahn Young-Ho;Lee Jin-Woo
    • Journal of Soil and Groundwater Environment
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    • v.10 no.2
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    • pp.52-58
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    • 2005
  • In Korea, little attention has been paid to microbial perchloroethylene (PCE) and/or trichloroethylene (TCE) dechlorination activity and identification of microorganisms involved in PCE reductive dechlorination at a PCE-contaminated aquifer. We performed microcosm tests using the groundwater samples from 4 different contaminated sites (i.e. Changwon A, Changwon B, Bucheon and Yangsan) to assess PCE reductive dechlorination activity. We also adapted molecular techniques to screen what types of known reductive dechlorinators are present at the PCE-contaminated aquifers. In the Changwon A and Changwon B active microcosms where potential electron donors such as sodium propionate, sodium lactate, sodium butyrate, and sodium fumarate, were added, ethylene, an end-product of complete reductive dechlorination of PCE, was detected after a period of 90 days of incubation. In the Bucheon and Yangsan active microcosms, cis-1,2-dichloroethylene (c-DCE) was accumulated without the production of vinyl chloride (VC) and ethylene. Molecular techniques were used to evaluate the microbial community structures in the Changwon B and Yangsan aquifer. We found two sequence types that were closely related to a known PCE to ethylene dechlorinator, named uncultured bacterium clone DCE47, in the Changwon B site clone library. However, in the Yangsan site clone library, no sequence type was closely related to known PCE dechlorinators reported. It is plausible that microorganisms being capable of completely dechlorinating PCE to ethylene may be present in the Changwon B site aquifer. In this study we find that complete PCE reductive dechlorinators are present at some PCE-contaminated sites in Korea. In an engineering point of view this information makes it feasible to apply a biological reductive dechlorination process for remediating PCE- and/or TCE-contaminated aquifers in Korea.

Assessment of Organic Compound and Bioassay in Soil Using Pharmaceutical Byproduct and Cosmetic Industry Wastewater Sludge as Raw Materials of Compost (제약업종 부산물 및 화장품 제조업 폐수처리오니 처리토양에 대한 유기화합물 및 Bioassay 분석 평가)

  • Lim, Dong-Kyu;Lee, Sang-Beom;Lee, Seung-Hwan;Nam, Jae-Jak;Na, Young-Eun;Kwon, Jang-Sik;Kwon, Soon-Ik;So, Kyu-Ho
    • Korean Journal of Environmental Agriculture
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    • v.23 no.4
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    • pp.203-210
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    • 2004
  • This study was conducted to assessment organic compound and bioassay (density of inhabited animal, fluctuation of predominant fungi, and survival ratio of earthworm) for finding damage on red pepper by heavily amount application of sludges in soil, which was treated with 3 pharmaceutical byproducts and a cosmetic industry wastewater sludge as raw materials of compost, and for establishing estimation method. HEM contents in the soil treated with pharmaceutical byproducts sludge2 (PS2) and cosmetic sludge (CS) were 0.51, 1.10 mg/kg respectively. PAHs content of PS2 treatment in the soil was 3406.8 ug/kg on July 8. In abundance of soil faunas, the pharmaceutical byproducts sludge2 treatment was the most highest. The next was decreased in the order of pig manure (PM) and the cosmetic sludge treatment. However the other pharmaceutical sludge treatments were remarkably reduced populations of soil inhabited animals. In upland soil treated with organic sludges, the numbers of bacteria and fungi of the pharmaceutical sludge treatment were 736, 909 cfu/g and those of the cosmetic sludge treatment were 440, 236 cfu/g, respectively. The pharmaceutical sludge treatments and the cosmetic sludge treatment in identification of predominant bacteria were not any tendency to compare with non fertilizer and pig manure treatments, but they had diverse bacteria than NPK treatment. In microcosm tests, the survival of the tiger earthworm in five soil samples was hardly affected against the soil of PSI (20%) after three months treated in the upland But after six months, survival of PS1 was 80%. At present, raw material of compost was authorized by contents of organic matter, heavy metal (8 elements), and product processing according to 'The specified gist on possible materials of using after analysis and investigation among raw materials of compost', however, for preparing to change regulation of raw material of compost and for considering to possibility of application, this study was conducted to investigate toxic organic compound and bioassay methods using inhabited animal, fungi, and earthworm without current regulation.

Field Studios of In-situ Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons

  • Semprini, Lewts
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.3-4
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    • 2004
  • Results will be presented from two field studies that evaluated the in-situ treatment of chlorinated aliphatic hydrocarbons (CAHs) using aerobic cometabolism. In the first study, a cometabolic air sparging (CAS) demonstration was conducted at McClellan Air Force Base (AFB), California, to treat chlorinated aliphatic hydrocarbons (CAHs) in groundwater using propane as the cometabolic substrate. A propane-biostimulated zone was sparged with a propane/air mixture and a control zone was sparged with air alone. Propane-utilizers were effectively stimulated in the saturated zone with repeated intermediate sparging of propane and air. Propane delivery, however, was not uniform, with propane mainly observed in down-gradient observation wells. Trichloroethene (TCE), cis-1, 2-dichloroethene (c-DCE), and dissolved oxygen (DO) concentration levels decreased in proportion with propane usage, with c-DCE decreasing more rapidly than TCE. The more rapid removal of c-DCE indicated biotransformation and not just physical removal by stripping. Propane utilization rates and rates of CAH removal slowed after three to four months of repeated propane additions, which coincided with tile depletion of nitrogen (as nitrate). Ammonia was then added to the propane/air mixture as a nitrogen source. After a six-month period between propane additions, rapid propane-utilization was observed. Nitrate was present due to groundwater flow into the treatment zone and/or by the oxidation of tile previously injected ammonia. In the propane-stimulated zone, c-DCE concentrations decreased below tile detection limit (1 $\mu$g/L), and TCE concentrations ranged from less than 5 $\mu$g/L to 30 $\mu$g/L, representing removals of 90 to 97%. In the air sparged control zone, TCE was removed at only two monitoring locations nearest the sparge-well, to concentrations of 15 $\mu$g/L and 60 $\mu$g/L. The responses indicate that stripping as well as biological treatment were responsible for the removal of contaminants in the biostimulated zone, with biostimulation enhancing removals to lower contaminant levels. As part of that study bacterial population shifts that occurred in the groundwater during CAS and air sparging control were evaluated by length heterogeneity polymerase chain reaction (LH-PCR) fragment analysis. The results showed that an organism(5) that had a fragment size of 385 base pairs (385 bp) was positively correlated with propane removal rates. The 385 bp fragment consisted of up to 83% of the total fragments in the analysis when propane removal rates peaked. A 16S rRNA clone library made from the bacteria sampled in propane sparged groundwater included clones of a TM7 division bacterium that had a 385bp LH-PCR fragment; no other bacterial species with this fragment size were detected. Both propane removal rates and the 385bp LH-PCR fragment decreased as nitrate levels in the groundwater decreased. In the second study the potential for bioaugmentation of a butane culture was evaluated in a series of field tests conducted at the Moffett Field Air Station in California. A butane-utilizing mixed culture that was effective in transforming 1, 1-dichloroethene (1, 1-DCE), 1, 1, 1-trichloroethane (1, 1, 1-TCA), and 1, 1-dichloroethane (1, 1-DCA) was added to the saturated zone at the test site. This mixture of contaminants was evaluated since they are often present as together as the result of 1, 1, 1-TCA contamination and the abiotic and biotic transformation of 1, 1, 1-TCA to 1, 1-DCE and 1, 1-DCA. Model simulations were performed prior to the initiation of the field study. The simulations were performed with a transport code that included processes for in-situ cometabolism, including microbial growth and decay, substrate and oxygen utilization, and the cometabolism of dual contaminants (1, 1-DCE and 1, 1, 1-TCA). Based on the results of detailed kinetic studies with the culture, cometabolic transformation kinetics were incorporated that butane mixed-inhibition on 1, 1-DCE and 1, 1, 1-TCA transformation, and competitive inhibition of 1, 1-DCE and 1, 1, 1-TCA on butane utilization. A transformation capacity term was also included in the model formation that results in cell loss due to contaminant transformation. Parameters for the model simulations were determined independently in kinetic studies with the butane-utilizing culture and through batch microcosm tests with groundwater and aquifer solids from the field test zone with the butane-utilizing culture added. In microcosm tests, the model simulated well the repetitive utilization of butane and cometabolism of 1.1, 1-TCA and 1, 1-DCE, as well as the transformation of 1, 1-DCE as it was repeatedly transformed at increased aqueous concentrations. Model simulations were then performed under the transport conditions of the field test to explore the effects of the bioaugmentation dose and the response of the system to tile biostimulation with alternating pulses of dissolved butane and oxygen in the presence of 1, 1-DCE (50 $\mu$g/L) and 1, 1, 1-TCA (250 $\mu$g/L). A uniform aquifer bioaugmentation dose of 0.5 mg/L of cells resulted in complete utilization of the butane 2-meters downgradient of the injection well within 200-hrs of bioaugmentation and butane addition. 1, 1-DCE was much more rapidly transformed than 1, 1, 1-TCA, and efficient 1, 1, 1-TCA removal occurred only after 1, 1-DCE and butane were decreased in concentration. The simulations demonstrated the strong inhibition of both 1, 1-DCE and butane on 1, 1, 1-TCA transformation, and the more rapid 1, 1-DCE transformation kinetics. Results of tile field demonstration indicated that bioaugmentation was successfully implemented; however it was difficult to maintain effective treatment for long periods of time (50 days or more). The demonstration showed that the bioaugmented experimental leg effectively transformed 1, 1-DCE and 1, 1-DCA, and was somewhat effective in transforming 1, 1, 1-TCA. The indigenous experimental leg treated in the same way as the bioaugmented leg was much less effective in treating the contaminant mixture. The best operating performance was achieved in the bioaugmented leg with about over 90%, 80%, 60 % removal for 1, 1-DCE, 1, 1-DCA, and 1, 1, 1-TCA, respectively. Molecular methods were used to track and enumerate the bioaugmented culture in the test zone. Real Time PCR analysis was used to on enumerate the bioaugmented culture. The results show higher numbers of the bioaugmented microorganisms were present in the treatment zone groundwater when the contaminants were being effective transformed. A decrease in these numbers was associated with a reduction in treatment performance. The results of the field tests indicated that although bioaugmentation can be successfully implemented, competition for the growth substrate (butane) by the indigenous microorganisms likely lead to the decrease in long-term performance.

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