• Title/Summary/Keyword: sequential incubation

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Desmutagenic Effect of Leonurus sibiricus L. to Aflatoxin B1 in Salmonella Mutation Assay (아플라톡신에 대한 익모초의 돌연변이 억제 효과)

  • 안병용;이갑상
    • The Korean Journal of Food And Nutrition
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    • v.9 no.3
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    • pp.294-298
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    • 1996
  • By the 505 chromotest which utilized Escherichia bolt PQ 37, Korean medicinal plants had been screened to Investigate the antimutagenic effect to aflatoxin B1(AFBl). Ikmocho(IMC, Leonurus sibiricus L.) was extracted with hot water. The extract was not found to be mutagenic in the Salmonella mutation test with or without metabolic activation, and the extract was showed to possess the antimutagenic properties towards AFB1-induced metation. The mutagenicity of AFB1 was inhibited by methanol soluble fracstion (IMC-MS) in dose-dependent. However, water-soluble fraction exhibited comutagenic activity. The greatest inhibitory effect of IMC-MS on AFB1 mutagenicity occurred when IMC-MS was first incubated, AFB1 followed by a second incubation with the cells and 59 mixture. Also lower inhibition was occurred when S9 mixtures were first incubated, with IMC-MS followed by a second incubation with AFBI. The results of the sequential incubation study support the probability that one mechanism of inhibition could involve the formation of chemical complex between IMC-MS and AFB1 rather than deactivation of S9 enzyme.

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Characteristics of Klebsiella pneumoniae exposed to serial antibiotic treatments (항생제 노출에 따른 Klebsiella pneumoniae의 내성 특성)

  • Jung, Lae-Seung;Jo, Ara;Kim, Jeongjin;Ahn, Juhee
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.428-436
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    • 2016
  • The emergence of antibiotic-resistant bacteria has been increased and become a public health concern worldwide. Many bacterial infections can be sequentially treated with different types of antibiotics. Thus, this study was designed to evaluate the changes in survival, antibiotic susceptibility, mutant frequency, ${\beta}$-lactamase activity, biofilm formation, and gene expression in Klebsiella pneumoniae after exposure to sequential antibiotic treatments of ciprofloxacin and meropenem. Treatments include control (CON; no addition), 1/2 MIC ciprofloxacin addition (1/2CIP), 2 MIC ciprofloxacin addition (2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC ciprofloxacin (16 h-incubation) (1/2CIP-1/2MER-2CIP), initial 1/2 MIC ciprofloxacin addition followed by 1/2 MIC meropenem (8 h-incubation) and 2 MIC meropenem (16 h-incubation) (1/2CIP-1/2MER-2MER), and initial 1/2 MIC ciprofloxacin addition followed by 2 MIC ciprofloxacin(8 h-incubation) and 2 MIC meropenem(16 h-incubation) (1/2CIP-2CIP-2MER). No growth of K. pneumoniae was observed for the 2CIP throughout the incubation period. The numbers of planktonic cells varied with the treatments (7~10 log CFU/ml), while those of biofilm cells were not significantly different among treatments after 24-h incubation, showing approximately 7 log CFU/ml. Among the sequential treatments, the least mutant frequency was observed at the 1/2CIP-1/2MER-2CIP (14%). Compared to the CON, 1/2CIP-2CIP-2MER decreased the sensitivity of K. pneumoniae to piperacillin, cefotaxime, and nalidixic acid. The highest ${\beta}$-lactamase activity was 22 nmol/min/ml for 1/2CIP-1/2MER-2CIP, while the least ${\beta}$-lactamase activity was 6 nmol/min/ml for 1/2CIP-2CIP-2MER. The relative expression levels of multidrug efflux pump-related genes (acrA, acrB, and ramA) were increased more than 2-fold in K. pneumoniae exposed to 1/2CIP-1/2MER-2MER and 1/2CIP-2CIP-2MER. The results suggest that the sequential antibiotic treatments could change the antibiotic resistance profiles in K. pneumoniae.

Remediation of As-contaminated Soil Using Magnetite and Bottom Ash (비소 오염 토양의 복원을 위한 자철석과 바닥재 활용)

  • Se Jin Oh;Min Woo Kang;Jong Cheol Lee;Hun Ho Lee;Hyun-Seog Roh;Yukwon Jeon;Dong Jin Kim;Sang Soo Lee
    • Korean Journal of Environmental Agriculture
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    • v.41 no.4
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    • pp.223-229
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    • 2022
  • BACKGROUND: Mining activities, smelter discharges, and sludges are the major sources of heavy metal contamination to soils. The objective of this study was to determine the efficiency of magnetite and bottom ash derived from coal ash in remediating As-contaminated soil. METHODS AND RESULTS: An incubation experiment was conducted for 10 weeks. Magnetite and bottom ash at different rates and ratios were applied to each plastic bottle repacked with 1,000 g of dried As-contaminated soil. After 3-weeks of incubation, the concentrations of available As were measured by using Mehlich-3, SBET, and sequential extraction methods. All of the subjected soil amendments resulted in significant decreases in available As concentration compared to the controls. The addition of magnetite at the highest rate was the best to stabilize As in the soils; however, the values of As concentration varied with the extraction methods. CONCLUSION(S): To ensure the stabilization accuracy of heavy metals in soil, both single and sequential extractions are recommended. The magnetite derived from fly coal ash can also be applicable as a heavy metal stabilizer for the As-contaminated soil.

Preferential Decomposition of Nitrogen during Early Diagenesis of Sedimentary Organic Matter (퇴적물 내 유기물의 초기 속성 작용에 나타난 유기 질소의 선택적 분해)

  • Han, Myung-Woo;Lee, Khang-Hyun
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.6 no.2
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    • pp.63-70
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    • 2001
  • Changes in concentrations of dissolved oxygen, ammonia, nitrate, pH, Fe and Mn were monitored from the laboratory incubation of an benthic chamber. The extent of sedimentary organic carbon and nitrogen decomposition was quantified by applying the concentration data to the chemical reaction equations of early diagenesis. The patterns of the concentration changes, observed during the 237 hr long incubation experiment, made it possible to divide the entire experiment period into four characteristic sub-periods (0-9 hr, 9-45 hr, 45-141hr, 141-237 hr). C/N ratio, estimated for each sub-period, was 6.63, 1.49, 0.81 and 0.02, respectively. This sequential decrease in C/N ratio suggests that during the incubation experiment dissolved nitrogen species diffuse more out of the sediment than dissolved carbon species. Greater diffusion of nitrogen indicates the preferential decomposition of organic nitrogen during early diagenesis of sedimentary organic matter. Comparison of the concentration data (sedimentary organic carbon and nitrogen, porewater organic carbon and ammonia)between the sediment pre and post incubation also indicates the preferential decomposition of nitrogen during early diagenesis of sedimentary organic matter.

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Bioprocess Development for Production of Alkaline Protease by Bacillus pseudofirmus Mn6 Through Statistical Experimental Designs

  • Abdel-Fattah, Y.R.;El-Enshasy, H.A.;Soliman, N.A.;El-Gendi, H.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.378-386
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    • 2009
  • A sequential optimization strategy, based on statistical experimental designs, is employed to enhance the production of alkaline protease by a Bacillus pseudofirmus local isolate. To screen the bioprocess parameters significantly influencing the alkaline protease activity, a 2-level Plackett-Burman design was applied. Among 15 variables tested, the pH, peptone, and incubation time were selected based on their high positive significant effect on the protease activity. A near-optimum medium formulation was then obtained that increased the protease yield by more than 5-fold. Thereafter, the response surface methodology(RSM) was adopted to acquire the best process conditions among the selected variables, where a 3-level Box-Behnken design was utilized to create a polynomial quadratic model correlating the relationship between the three variables and the protease activity. The optimal combination of the major medium constituents for alkaline protease production, evaluated using the nonlinear optimization algorithm of EXCEL-Solver, was as follows: pH of 9.5, 2% peptone, and incubation time of 60 h. The predicted optimum alkaline protease activity was 3,213 U/ml/min, which was 6.4 times the activity with the basal medium.

Development of FK506-hyperproducing strain and optimization of culture conditions in solid-state fermentation for the hyper-production of FK506

  • Mo, SangJoon;Yang, Hyeong Seok
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.289-298
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    • 2016
  • FK506 hyper-yielding mutant, called the TCM8594 strain, was made from Streptomyces tsukubaensis NRRL 18488 by mutagenesis using N-methyl-N'-nitro-N-nitrosoguanidine, ultraviolet irradiation, and FK506 sequential resistance selection. FK506 production by the TCM8594 strain improved 45.1-fold ($505.4{\mu}g/mL$) compared to that of S. tsukubaensis NRRL 18488 ($11.2{\mu}g/mL$). Among the five substrates, wheat bran was selected as the best solid substrate to produce optimum quantities of FK506 ($382.7{\mu}g/g$ substrate) under solid-state fermentation, and the process parameters affecting FK506 production were optimized. Maximum FK506 yield ($897.4{\mu}g/g$ substrate) was achieved by optimizing process parameters, such as wheat bran with 5 % (w/w) dextrin and yeast extract as additional nutrients, 70 % (v/w) initial solid substrate moisture content, initial medium pH of 7.2, $30^{\circ}C$ incubation temperature, inoculum level that was 10 % (v/w) of the cell mass equivalent, and a 10 day incubation. The results showed an overall 234 % increase in FK506 production after optimizing the process parameters.

Application of Zeolite with Different Cation Exchange Capacity for the Stabilization of Heavy Metals in Upland Soil (양이온교환용량이 다른 제올라이트 처리에 따른 밭토양 내 중금속 안정화 평가)

  • Gu, Bon-Wun;Kim, Mun-Ju;Park, Seong-Jik
    • Journal of The Korean Society of Agricultural Engineers
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    • v.59 no.5
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    • pp.41-49
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    • 2017
  • This study was aimed to investigate the influence of cation exchange capacity (CEC) and application amounts of zeolite on the stabilization of heavy metals (As, Ni, Pb, and Zn) in upland soils. The upland soils were sampled from field near mines located in Gyeonggi Province. The CEC of zeolite was treated at three different levels, ie, low, medium, and high, while zeolite was amended with soils at the ratio of 0.1 % and 0.5 % as to soil weight. A sequential extraction was performed for the soil sampled at 1, 2 4, and 8 week after zeolite was added to the soil. The concentrations of Pb and Zn appeared to be high in the sampled soils. The mobility of heavy metals obtained from sequential experiments was as follows: Pb > Zn > Ni >As. Addition of zeolite to contaminated soils effectively reduced exchangeable and carbonate fractions but increased organic and residual fraction, indicating that zeolite is effective for immobilizing heavy metals in soils. The influence of incubation time on the metal stabilization was rather pronounced as compared to the application amount and CEC of zeolite.

A Study on the Decomposition of Amygdalin Using an In Vitro Assay (Amygdalin의 in Vitro 분해에 관한 연구)

  • Kwon, Hoon-Jeong;Jo, Yong-Jin
    • Toxicological Research
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    • v.23 no.1
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    • pp.47-53
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    • 2007
  • Amygdalin is a cyanogenic glycoside which is commonly found in almonds, bamboo shoots, and apri-cot kernels, and peach kernels. Amygdalin was first hydrolysed into prunasin, then degraded into cyanohydrin by sequential two-stage mechanism. The objective of this study was to examine the amygdalin decomposition and cyanide formation at various in vitro conditions, including acid, enzyme and anaerobic microbes (AM) in human feces (HF). In acid hydrolysis mimicking gastric environment, amygdalin was degraded to cyanide up to 0.2% in specific pH. In contrast, enzyme assay showed higher cyanide generation either by ${\beta}$-glucosidase, or by incubation with microbe. In conclusion, we are convinced of cyanide generation are occurred mainly by microbiological activities of the gut flora up to 41.53%. After ingestion with some staff, the degree and site of degradation in an organism is a key parst of regulatory decision making of that staff.

Biodegradation of Hydrocarbon Contamination by Immobilized Bacterial Cells

  • Rahman Raja Noor Zaliha Abd.;Ghazali Farinazleen Mohamad;Salleh Abu Bakar;Basri Mahiran
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.354-359
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    • 2006
  • This study examined the capacity of immobilized bacteria to degrade petroleum hydrocarbons. A mixture of hydrocarbon-degrading bacterial strains was immobilized in alginate and incubated in crude oil-contaminated artificial seawater (ASW). Analysis of hydrocarbon residues following a 30-day incubation period demonstrated that the biodegradation capacity of the microorganisms was not compromised by the immobilization. Removal of n-alkanes was similar in immobilized cells and control cells. To test reusability, the immobilized bacteria were incubated for sequential increments of 30 days. No decline in biodegradation capacity of the immobilized consortium of bacterial cells was noted over its repeated use. We conclude that immobilized hydrocarbon-degrading bacteria represent a promising application in the bioremediation of hydrocarbon-contaminated areas.

Biodegradation of Pentachlorophenol by Various White Rot Fungi (수질분해균(水質分解菌)에 의한 Pentachlorophenol의 미생물분해(微生物分解))

  • Choi, In-Gyu;Ahn, Sye-Hee
    • Journal of the Korean Wood Science and Technology
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    • v.26 no.3
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    • pp.53-62
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    • 1998
  • In this research, 7 species of white rot fungi were used for determining the resistance against pentachlorophenol (PCP). Three fungi with good PCP resistance were selected for evaluating the biodegradability, and biodegradation mechanism by HPLC and GC/MS spectrometry. Among 7 fungi, there were significant differences on PCP resistance on 4 different PCP concentrations. In the concentrations of 50 and 100ppm ($\mu$g of PCP per g of 2% malt extract agar), most fungi were easily able to grow, and well suited to newly PCP-added condition, but in that of more than 250ppm, the mycelia growths of Ganoderma lucidum 20435, G. lucidum 20432, Pleurotus ostreatus, and Daldinia concentrica were significantly inhibited or even stopped by the addition of PCP to the culture. However, Trametes versicolor, Phanerochaete chrysosporium, and Inonotus cuticularis still kept growing at 250ppm, indicating the potential utilization of wood rot fungi to high concentrated PCP biodegradation. Particularly, P. chrysosporium even showed very rapid growth rate at more than 500ppm of PCP concentration. Three selected fungi based on the above results showed an excellent biodegradability against PCP. P. chrysosporium degraded PCP up to 84% on the first day of incubation, and during 7 days, most of added PCP were degraded. T. versicolor also showed more than 90% of biodegradability at 7th day, and even though the initial stage of degradation was very slow, I. cuticularis has been approached to 90% at 21 st day after incubation with dense growing pattern of mycelia. Therefore, the PCP biodegradability was definitely dependent on the rapid suitability of fungi to newly PCP-added condition. In addition, the PCP biodegradation by filtrates of P. chrysosporium, T. versicolor, and I. cuticularis was very minimal or limited, suggesting that the extracellular enzyme system may be not so significantly related to the PCP biodegradation. Among the biodegradation metabolites of PCP, the most abundant one was pentachloroanisole which resulted in a little weaker toxicity than PCP, and others were tetrachlorophenol, tetrachloro-hydroquinone, benzoic acid, and salicylic acid, suggesting that PCP may be biodegraded by several sequential reactions such as methylation, radical-induced oxidation, dechlorination, and hydroxylation.

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