• Title/Summary/Keyword: Microbial detection

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Evaluation of Natural Antimicrobial Formulas against Yeasts in Functional Bevereges (기능성 음료 중 효모 제어를 위한 천연항균제 혼합조성 물 평가)

  • Yeon, Ji-Hye;Jo, Seo-Hee;Lee, Hee-Seok;Ha, Sang-Do;Park, Chul-Soo;Woo, Moon-Jea;Lee, Sang-Hun;Kim, Jin-Soo;Lee, Chon
    • Journal of Food Hygiene and Safety
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    • v.24 no.3
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    • pp.189-193
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    • 2009
  • The purpose of this study was to investigate the antimicrobial activities of natural antimicrobials (10 formulas, $F1{\sim}F10$) against yeasts in functional beverages. The growth rates of yeasts were different with the ten different natural antimicrobial formulas tested. Yeasts grew for 14 days and the antimicrobial effect was observed between 14 and 18 days. Levels of S. cerevisiae, Z. bailii, and P. membranaefaciens were reduced to the limit of detection (ND) < 10 CFU/mL) after 28 days. Resistance against the antimicrobial effect was greatest for P. membranaefaciens, which grew to a level of $0.12{\sim}1.48\;\log_{10}\;CFU$/mL after 14 days and was reduced to a level of $1.61{\sim}3.55\;\log_{10}\;CFU$/mL after 28 days. The resistance of C. albicans was also high with a growth level of $0.13{\sim}1.28\;\log_{10}\;CFU$/mL after 14 days and reduction to $1.51{\sim}5.30\;\log_{10}\;CFU$/mL after 28 days. The antimicrobial effect of F10 was strongest for P. membranaefaciens. Every treatment reduced the microbial levels to $2.68{\sim}5.62\;\log_{10}\;CFU$/mL after 6 months. F2, F4, F5, F6, and F10 reduced the C. albicans level to ND after 28 days while F1, F3, F8, and F9 reduced yeasts to the ND level after 6 months. The antimicrobial activities observed here will be useful for development of natural antimicrobials.

Characteristics of Histamine Forming Bacteria from Tuna Fish Waste in Korea (국내 참치 부산물 내 히스타민 생성 주요 세균의 특성 구명)

  • Bang, Min-Woo;Chung, Chang-Dae;Kim, Seon-Ho;Chang, Moon-Baek;Lee, Sung-Sil;Lee, Sang-Suk
    • Journal of Life Science
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    • v.19 no.2
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    • pp.277-283
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    • 2009
  • Biogenic amines are generally formed through the decarboxylation of specific free amino acids by exogenous decarboxylases released by microbial species associated with the fish products and fermented feeds. This study was conducted to investigate the properties of e tuna waste regarding the control of degradation of biogenic amines (histamine, tyramine, tryptamine, putrescine, and cadaverine) that might be related with the anti-nutritional factor of the tuna waste that is used for manufacturing domestic fish meal. The values of pH and the salt content were 6.51, 3.35% in tuna waste and 5.58 and 5.83% in tuna fish meal, respectively. The strains and dominant bacteria tested in the tuna waste sample were 9.20, 9.29, 5.67, 7.82 and 7.58 log CFU/g of total bacteria, aerobic plate count (APC), total coliform (TC), Lactobacillus spp. and Bacillus spp., respectively. The main histamine forming-bacteria (HFB) in tuna waste were detected by silica gel thin-layer chromatography (TLC) and 7 histamine-forming bacterial species were isolated among microbes grown in selective medium. The histamine concentration was determined by detection of fluorescence of ο-phthaldialdehyde (OPA) derivatives using HPLC and the date were used to reconfirm the identities of the amine-producing bacteria. The 15 histamine- forming bacteria strains grown in trypicase soy broth (TSB) supplemented with 1% L-histidine (TSBH) were identified as Lactococcus(L.) lactis subsp. lactis, Klebsiella pneummonlae, L. garvieae 36, Vibrio olivaceus, Hafnia alvei and L. garvieae which were main dominant amine - producing strains, and Morganella morganii identified by 16S ribosomal RNA (rRNA) sequencing with PCR amplification. A Phylogenetic tree generated from the 16S rRNA sequencing data showed different phyletic lines that could be readily classified as biogenic amine forming gram-positive and negative bacteria.

Effects of Glufosinate-Ammonium to Earthworms, Soil Microorganisms and Crops (제초제 glufosinate-ammonium의 지렁이 및 토양 미생물과 작물에 미치는 영향)

  • Kim, Yong-Seog;Jeon, Yong-Bae;Choi, Hae-Jin;Kim, Song-Mun;Kim, Sung-Min
    • The Korean Journal of Pesticide Science
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    • v.10 no.2
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    • pp.76-83
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    • 2006
  • In order to investigate the impacts of non-selective herbicide, glufosinate-ammonium (ammonium 4-[hydroxy(methyl)phosphinoyl] -DL-homoalaninate, GLA) to the non-target organisms, earthworm was exposed to GLA in the field soil for a month, and microbial populations in the soil were investigated after application of GLA. Simultaneously, the residues of GLA and its metabolite, 3-MPP were analyzed in the same soil. Meanwhile, to elucidate the influence of GLA to the growth of non-target crops incase of inter-furrow application, the amounts of carotenoid, chlorophyll, amino acid, proteins and sugars in the leaves of potato and chinese cabbage grown in the same field were investigated. In result, the dead earthworm was not observed during the test period, and the increasing rates of bodyweight were $9.410{\sim}11.603%$ in GLA-treated plots and 5.645% in GLA-untreated plots. The populations of fungi, bacteria and actinomycetes in the GLA-treated soils were $6.2{\times}10^4$, $1.5{\times}10^6$ and $5.7{\times}10^4$, respectively. They maintained relatively similar levels to the control which were $3.7{\times}10^4$, $3.7{\times}10^5$ and $3.7{\times}10^4$, respectively. In residue analysis, the limit of detection of GLA was 0.02 mg $kg^{-1}$, that of 3-MPP was the same level, and the half-life of GLA was 15 days in sandy clay loam soil. This result indicates that GLA was degraded very quickly in field soil. On the other hand, the amounts of physiological, biochemical components such as carotenoid, amino acid, chlorophyll, protein and sugar were ranged from 90.0 to 104.3% in potato and from 99.0 to 112.7% in chinese cabbage. Comparing with hand-weeded plots, it is indicated that GLA had not affected to the growth of non-target crops when applied at inter-furrow in crops-growing field.

Mobility of Nitrate and Phosphate through Small Lysimeter with Three Physico-chemically Different Soils (소형 라이시메터시험을 통한 토양특성에 따른 질산과 인산의 이동성 비교)

  • Han, Kyung-Hwa;Ro, Hee-Myong;Cho, Hyun-Jun;Kim, Lee-Yul;Hwang, Seon-Woong;Cho, Hee-Rae;Song, Kwan-Cheol
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.4
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    • pp.260-266
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    • 2008
  • Small lysimeter experiment under rain shelter plastic film house was conducted to investigate the effect of soil characteristics on the leaching and soil solution concentration of nitrate and phosphate. Three soils were obtained from different agricultural sites of Korea: Soil A (mesic family of Typic Dystrudepts), Soil B (mixed, mesic family of Typic Udifluvents), and Soil C (artificially disturbed soils under greenhouse). Organic-C contents were in the order of Soil C ($32.4g\;kg^{-1}$) > Soil B ($15.0g\;kg^{-1}$) > Soil A ($8.1g\;kg^{-1}$). Inorganic-N concentration also differed significantly among soils, decreasing in the order of Soil B > Soil C > Soil A. Degree of P saturation (DPS) of Soil C was 178%, about three and fifteen times of Soil B (38%) and Soil A (6%). Prior to treatment, soils in lysimeters (dia. 300 mm, soil length 450 mm) were tabilized by repeated drying and wetting procedures for two weeks. After urea at $150kg\;N\;ha^{-1}$ and $KH_2PO_4$ at $100kg\;P_2O_5\;ha^{-1}$ were applied on the surface of each soil, total volume of irrigation was 213 mm at seven occasions for 65 days. At 13, 25, 35, 37, and 65 days after treatment, soil solution was sampled using rhizosampler at 10, 20, and 30 cm depth and leachate was sampled by free drain out of lysimeter. The volume of leachate was the highest in Soil C, and followed by the order of Soils A and B, whereas the amount of leached nitrate had a reverse trend, i.e. Soil B > Soil A > Soil C. Soil A and B had a significant increase of the nitrate concentration of soil solution at depth of 10 cm after urea-N treatment, but Soil C did not. High nitrate mobility of Soil B, compared to other soils, is presumably due to relatively high clay content, which could induce high extraction of nitrate of soil matrix by anion exclusion effect and slow rate of water flow. Contrary to Soil B, high organic matter content of Soil C could be responsible for its low mobility of nitrate, inducing preferential flow by water-repellency and rapid immobilization of nitrate by a microbial community. Leached phosphate was detected in Soil C only, and continuously increased with increasing amount of leachate. The phosphate concentration of soil solution in Soil B was much lower than in Soil C, and Soil A was below detection limit ($0.01mg\;L^{-1}$), overall similar to the order of degree of P saturation of soils. Phosphate mobility, therefore, could be largely influenced by degree of P saturation of soils but connect with apparent leaching loss only more than any threshold of P accumulation.

The Effect of Fermented Diet with Whole Crop Barley Silage on Fecal Shape and Odorous Compound Concentration from Feces in Pregnant Sows (청보리 사일리지 첨가 발효사료가 임신돈의 분 성상 및 악취물질 농도에 미치는 효과)

  • Cho, S.B.;Kim, C.H.;Hwang, O.H.;Park, J.C.;Kim, D.W.;Sung, H.G.;Yang, S.H.;Park, K.H.;Choi, D.Y.;Yoo, Y.H.
    • Journal of Animal Environmental Science
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    • v.17 no.3
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    • pp.145-154
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    • 2011
  • The study was conducted to investigate the effect of feeding fermented diet including whole crop barley silage on the odor reduction and microbial population change in feces, and the constipation prevention of pregnant sows. The concentration of phenol was not detected in tapioca, beet pulp, wheat bran and lupin seeds, while that of p-cresol was ranged between 9.62 and 52.11 mg/L showing that lupin was highest and tapioca was lowest. It was determined that tapioca and beet pulp were useful feed ingredients to reduce odor due to their lower contents of phenol and indole compounds. Ten pregnant sows were allocated to control group and fermented diet group in 5 sows in each group. They were fed 3.0 kg DM/d of diets for 28 days. Feces was examined and showed that the feces from the fermented diet group was observed with the higher moisture content and the lower hardness than that of the control diet group and the population of E. coli was decreased and the population of lactobacilus was higher than that of the control diet group. The concentrations of p-cresol and skatole were lower than the detection levels at 33% and 67% among the samples of feces of the control group and at 67% and 100% among the samples of the feces of fermented diet group respectively. Thus it is expected that the odor from the feces of pregnant sows fed the fermented diet could be reduced compared with that of control group. Therefore, it is suggested that feeding fermented whole barley diet to pregnant sows improve the function of intestine and reduce the rate of occurrences of constipation and odor levels.

Antimicrobial, antifungal effect and safety verification using BCOP assay of extracts from Coptis chinensis (황련(Coptis chinensis) 추출물의 항균, 항진균 효과와 BCOP 분석을 이용한 안전성 검증)

  • Kim, Eun-Hee;Jang, Young-Ah;Kim, Sol-Bi;Kim, Han-Hyuk;Lee, Jin-Tae
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.297-304
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    • 2018
  • Coptis chinensis is used in oriental medicine for soothing, anti-inflammation, antimicrobial and antipyretic properties, and its main ingredient berberine is known to have strong antibacterial activity. In this study, we investigated the anti-microbial effect of hot water extract of Coptis chinensis (CW) on skin related microorganism and the airborne microbe, the antifungal effects of fungi, which are frequently detected in residential environments. CW showed antibacterial effect against Propionibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis, against the airborne microbe, which was collected in four different places. At the concentration of 100 mg/mL, the antimicrobial activity continued for 42 days, showed heat stability without change in the antimicrobial activity even after heat treatment. The MIC and MBC of CW against S. aureus was 0.03, 0.05 mg/mL, against S. epidermidis was 0.50, 0.75 mg/mL and against P. acne was 0.10, 0.15 mg/mL. As a result of measuring the MIC of four kinds of fungi with high detection frequency in the surrounding environment, Gliocladium virens was 65 mg/mL by determined as MIC which can inhibit one hundred percent of mycelial growth. The concentration 90 mg/mL was determined as MIC against Aureobasidium pullulans and 100 mg/mL against Penicilium pinophilum and Chaetomium globosum. CW was considered a safe extract that showed no irritation even in the ocular mucous membrane irritation evaluation test, a patch test. Therefore, these results suggest that Coptis chinensis has antimicrobial, antifungal and safety on human body and can be applied to the development of materials for cosmetic and residential environment industries.

Application of Polymerase Chain Reaction (PCR) to the Diagnosis of Tuberculosis (Polymerase Chain Reaction (PCR)을 이용한 결핵의 진단에 관한 연구)

  • Kim, Ho-Joong;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.6
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    • pp.517-525
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    • 1992
  • Background: Since its development by Saiki et al, polymerase chain reaction (PCR) has been very useful in various fields of molecular biology. PCR can be used for the detection of a very small amount of microbial agent, and is especially useful in those patients who are difficult to diagnose microbiologically or serologically. Mycobacterium tuberculosis is a very slowly growing organism and AFB staining frequently shows false negative results, and therefore PCR would be a very rapid, easy, and sensitive diagnostic method for the diagnosis of Mycobacterium tuberculosis. Method: To compare PCR with conventional methods in diagnosing Mycobacterium tuberculosis in sputum, we used sputa of patients who visited or were admitted to Seoul National University Hospital. The amplification targets were 383 base pair DNA, a part of 2520 base pair DNA encoding 65 kD Mycobacterium tuberculosis specific protein (the primers are TB-1, -2), and 123 base pair DNA, a part of IS6110 fragment, which multiple copies are known to exsist PCR one genome (the primers are Sal I-1, -2). We also requested AFB staing and culture to the lab of Seoul National University Hospital with the same sample and compared the results. Results: 1) Using TB-1, -2 primers, PCR was positive in 73.1% (19/26) of culture positive sputa, in 12.5% (1/8) of culture negative. but clinically diagnosed tuberculous sputa, and was negative in all sputa of patients who were clinically diagnosed as non-tuberculous etiology. 2) Using Sal I-I, -2 primers, PCR was positive in 94.1% (32/34) of culture positive sputa, in 23.1% (6/26) of culture negative, but clinically diagnosed tuberculous sputa, and was negative in 87.5% (14/16) of sputa from patients who were clinically diagnosed as non-tuberculous etiology. Conclusion: PCR could be a very rapid, sensitive and specific method for the diagnosis of Mycobacterium tuberculosis in sputa, and further studies should be followed for the development of easier method.

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High Hydrostatic Pressure Sterilization of Vibrio parahaemolyticus and Escherichia coli in Raw Oyster (생굴 중 Vibrio parahaemolyticus와 Escherichia coli의 초고압 살균)

  • Park, Whan-Jun;Jwa, Mi-Kyung;Hyun, Sun-Hee;Lim, Sang-Bin;Song, Dae-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.7
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    • pp.935-939
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    • 2006
  • Raw oyster (Crassostrea gigas) was inoculated with Vibrio parahaemolyticus and Escherichia coli, treated with high hydrostatic pressure and evaluated for microbial counts. Cell death of V. parahaemolyticus (Vp) increased with the increase of applied pressure. Vp starting inoculum of $3.8{\times}10^5\;CFU/mL$ was totally eliminated after exposure to 200 MPa for 10 min at $22^{\circ}C$ Viable cell of Vp decreased with the increase in treatment time and dropped below the detection limit with treament of 25 min at $22^{\circ}C/150\;MPa$. The number of Vp by treatment of $0^{\circ}C$ and $10^{\circ}C$ for 20 and 25 mon at 100 MPa, respectively. For E. coli, there was an initial lag up to 250 MPa gollowed by a rapid decline. Treatment at 325 MPa/$22^{\circ}C$ for 15 min caused 5-log reduction, while that at 375 MPa resulted in total reduction of starting inoculum of $4.0{\times}10^7\;CFU/mL$. Lower treatment temperature showed higher killing effect of E. coli at the same treatment pressure and time. Viable cell of E. coli decreased with the increase in treatment time, and 4-log reduction was achieved with treatment of 5 min at $10^{\circ}C$/350 MPa and then total reduction was achieved after treatment of 15 mon. Higher pressure, lower temperature and longer time were more effective in sterilizing V. parahaemolyticus and E. coli.

Characterization of compounds and quantitative analysis of oleuropein in commercial olive leaf extracts (상업용 올리브 잎 추출물의 화합물 특성과 이들의 oleuropein 함량 비교분석)

  • Park, Mi Hyeon;Kim, Doo-Young;Arbianto, Alfan Danny;Kim, Jung-Hee;Lee, Seong Mi;Ryu, Hyung Won;Oh, Sei-Ryang
    • Journal of Applied Biological Chemistry
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    • v.64 no.2
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    • pp.113-119
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    • 2021
  • Olive (Olea europaea L.) leaves, a raw material for health functional foods and cosmetics have abundant polyphenols including oleuropein (major bioactive compound) with various biological activities: antioxidant, antibacterial, antiviral, anticancer activity, and inhibit platelet activation. Oleuropein has been reported as skin protectant, antioxidant, anti-ageing, anti-cancer, anti-inflammation, anti-atherogenic, anti-viral, and anti-microbial activity. Despite oleuropein is the important compound in olive leaves, there is still no quantitative approach to reveal oleuropein content in commercial products. Therefore, a validated method of analysis has to develop for oleuropein. In this study, the components and oleuropein content in 10 types of products were analyzed using a developed method with ultra-performance liquid chromatography to quadrupole time-of-flight mass spectrometry, charge of aerosol detector, and photodiode array. The total of 18 compounds including iridoids (1, 3, 4, 14, and 16-18), coumarin (2), phenylethanoids (5, 9, and 11), flavonoids (6-8, 10, 12, and 13), lignan (15), were tentatively identified in the leaves extract based high resolution mass spectrometry data, and the content of oleuropein in each product was almost identical between two detection methods. The oleuropein in three commercial product (A, G, H) was contained more over the suggested content, and it of five products (B, E, H, I, J) were analyzed within 5-10% error range. However, the two products (C, D) were found far lower than suggested contents. This study provides that analytical results of oleuropein could be a potential information for the quality control of leaf extract for a manufactured functional food.

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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