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

  • Semprini, Lewts
    • 한국지하수토양환경학회:학술대회논문집
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    • 한국지하수토양환경학회 2004년도 총회 및 춘계학술발표회
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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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고구마 끝순 추출물의 항균 및 항돌연변이 효과 (Antibacterial and Antimutagenic Effects of Sweetpotato Tips Extract)

  • 이준설;신미진;박양균;안영섭;정미남;김학신;김정목
    • 한국작물학회지
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    • 제52권3호
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    • pp.303-310
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    • 2007
  • 고구마 용도 다양화와 지상부의 채소적 가치를 구명키 위하여, 괴근에서 자라난 싹 중 부드러운 잎과 잎자루를 포함한 줄기끝 10 cm의 끝순을 수확하여, 끝순 추출물의 페놀화합물 함량, 식중독균인 Listeria monocytogenes, Salmonella Typhimurium에 대한 항균 활성, 식품안전성과 관련된 돌연변이원성 및 항돌연변이원성 등을 조사한 결과는 다음과 같다. 1. 고구마 끝순에는 6종의 페놀화합물이 함유되어 있었으며, 그 중 chlorogenic acid와 caffeic acid의 함량이 총페놀함량의 $89{\sim}93%$로 대부분을 차지하였다. 또한 용매분획별 페놀화합물 함량은 ethyl acetate 및 buthyl alcohol 추출물이 다른 용매추출물에 비하여 월등히 높았다. 2. 고구마 끝순의 80% ethanol 조추출물의 수율은 12.5 g이었고, 용매분획별로는 water 분획물이 8.9 g으로 가장 많았으며, 다음은 chloroform 분획물이 1.6 g이었고, 나머지 용매 분획물은 $0.7{\sim}1.1g$의 수율을 나타내었다. 3. 고구마 끝순에는 6종의 페놀화합물이 함유되어 있었으며, 그 중 chlorogenic acid와 caffeic acid의 함량이 총페놀 함량의 $89{\sim}93%$로 대부분을 차지하였다. 또한 용매분획별 페놀화합물 함량은 ethyl acetate 및 buthyl alcohol 추출물이 다른 용매추출물에 비하여 월등히 높았다. 4. Listeria monocytogenes에 ethyl acetate 및 buthyl alcohol 추출물을 첨가한 결과 16시간까지는 3 log 이상 균의 성장을 억제하였으며 특히 20 mg/ml butanol 추출물은 균의 성장을 억제시키다 12시간 이후에는 완전히 균을 사멸시키는 것으로 나타났다. Salmonella Typhimurium에 대해서도 고구마 끝순 추출물은 $0.5{\sim}1$ log 정도의 감소를 나타내었다. 5. 고구마 끝숱 추출물의 Salmonella Typhimurium TA98 및 TA100에 대한 돌연변이 활성을 조사한 결과 TA98은 $29{\sim}33$ CFU/plate이었고, TA100은 $159{\sim}188$ CFU/plate로 돌연변이 활성이 없었다. 6. 직접 돌연변이원 물질인 2-NF와 MMS를 처리하여 복귀돌연변이수를 측정한 결과 ethyl acetate 및 buthyl alcohol에서 돌연변이로 인한 복귀변이원성 집락수가 낮게 나타남으로써 고구마 끝순 추출물이 항돌연변이 활성이 있는 것으로 나타났다.