• Proceedings of the Korean Environmental Sciences Society Conference
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• 1998.10a
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• pp.2-4
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• 1998

Proliferation of Nocardia amarae cells in activated sludge has often been associated with the generation of nuisance foams. Despite intense research activities in recent years to examine the causes and control of Nocardia foaming in activated sludge, the foaming continued to persist throughout the activated sludge treatment plants in United States. In addition to causing various operational problems to treatment processes, the presence of Nocardia may have secondary effects on the fate of heavy metals that are not well known. For example, for treatment plants facing more stringent metal removal requirements, potential metal removal by Nocardia cells in foaming activated sludge would be a welcome secondary effect. In contrast, with new viosolid disposal regulations in place (Code o( Federal Regulation No. 503), higher concentration of metals in biosolids from foaming activated sludge could create management problems. The goal of this research was to investigate the metal sorption property of Nocardia amarae cells grown in batch reactors and in chemostat reactors. Specific surface area and metal sorption characteristics of N. amarae cells harvested at various growth stages were compared. Three metals examined in this study were copper, cadmium and nickel. Nocardia amarae strain (SRWTP isolate) used in this study was obtained from the University of California at Berkeley. The pure culture was grown in 4L batch reactor containing mineral salt medium with sodium acetate as the sole carbon source. In order to quantify the sorption of heavy metal ions to N amarae cell surfaces, cells from the batch reactor were harvested, washed, and suspended in 30mL centrifuge tubes. Metal sorption studies were conducted at pH 7.0 and ionlc strength of 10-2M. The sorption Isotherm showed that the cells harvested from the stationary and endogenous growth phase exhibited significantly higher metal sorption capacity than the cells from the exponential phase. The sequence of preferential uptake of metals by N. amarae cells was Cu>Cd>Ni. The specific surFace area of Nocardia cells was determined by a dye adsorption method. N.amarae cells growing at ewponential phase had significantly less specific surface area than that of stationary phase, indicating that the lower metal sorption capacity of Nocardia cells growing at exponential phase may be due to the lower specific surface area. The growth conditions of Nocardia cells in continuous culture affect their cell surface properties, thereby governing the adsorption capacity of heavy metal. The comparison of dye sorption isotherms for Nocardia cells growing at various growth rates revealed that the cell surface area increased with increasing sludge age, indicating that the cell surface area is highly dependent on the steady-state growth rate. The highest specific surface area of 199m21g was obtained from N.amarae cell harvested at 0.33 day-1 of growth rate. This result suggests that growth condition not only alters the structure of Nocardia cell wall but also affects the surface area, thus yielding more binding sites of metal removal. After reaching the steady-state condition at dilution rate, metal adsorption isotherms were used to determine the equilibrium distributions of metals between aqueous and Nocardia cell surfaces. The metal sorption capacity of Nocardia biomass harvested from 0.33 day-1 of growth rate was significantly higher than that of cells harvested from 0.5- and 1-day-1 operation, indicatng that N.amarae cells with a lower growth rate have higher sorpion capacity. This result was in close agreement with the trend observed from the batch study. To evaluate the effect of Nocardia cells on the metal binding capacity of activated sludge, specific surface area and metal sorption capacity of the mixture of Nocardia pure cultures and activated sludge biomass were determined by a series of batch experiments. The higher levels of Nocardia cells in the Nocardia-activated sludge samples resulted in the higher specific surface area, explaining the higher metal sorption sites by the mixed luquor samples containing greater amounts on Nocardia cells. The effect of Nocardia cells on the metal sorption capacity of activated sludge was evaluated by spiking an activated sludge sample with various amounts of pre culture Nocardia cells. The results of the Langmuir isotherm model fitted to the metal sorption by various mixtures of Nocardia and activated sludge indicated that the mixture containing higher Nocardia levels had higher metal adsorption capacity than the mixture containing lower Nocardia levels. At Nocardia levels above 100mg/g VSS, the metal sorption capacity of activate sludge increased proportionally with the amount of Noeardia cells present in the mixed liquor, indicating that the presence of Nocardia may increase the viosorption capacity of activated sludge.

• Journal of Life Science
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• v.18 no.6
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• pp.789-795
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• 2008

To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and $30^{\circ}C$, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as $Mg^{+2}$, $Mn^{+2}$, but was inhibited by the addition of $Co^{+2}$, $Hg^{+2}$, $Zn^{+2}$ and $Ag^{+2}$. NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.

• Journal of Life Science
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• v.18 no.12
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• pp.1625-1630
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• 2008

A microorganism capable of producing high level of poly-3-hydoxybutyrate (PHB) from xylose was isolated from soil. The isolated strain J-65 was identified as Bacillus megaterium based on the morphological, biochemical and molecular biological characteristics. The optimum temperature and pH for the growth of B. megaterium J-65 were $37^{\circ}C$ and 8.0, respectively. The optimum medium composition for the cell growth was 2% xylose, 0.25% $(NH_4)_2SO_4$, 0.3% $Na_2HPO_4{\cdot}12H_2O$, and 0.1% $KH_2PO_4$. The optimum condition for PHB accumulation was same to the optimum condition for cell growth. Copolymer of ${\beta}$-hydroxybutyric and ${\beta}$-hydroxyvaleric acid was produced when propionic acid was added to shake flasks containing 20 g/l of xylose. Fermenter culture was carried out to produce the high concentration of PHB. In batch culture, cell mass was 9.82 g/l and PHB content was 35% of dry cell weight. PHB produced by B. megaterium J-65 was identified as homopolymer of 3-hydoxybutyric acid by GC and NMR.

• Journal of Life Science
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• v.24 no.4
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• pp.352-360
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• 2014

We evaluated the growth performance, biochemical characteristics, and immune responses in weaning pigs given a diet containing MR-1 (0.2%/feed) or antibiotics (0.1%/feed) for 45 days. In vitro study showed that MR-1 has antibacterial activity against a variety of strains of pathogenic bacteria, especially a strain of cattle-derived Escherichia coli K99 (E. coli K99) by agar diffusion assay. In the in vivo model, 0.2% MR-1-given group clearly ameliorated the weight gain and feed efficiency in the growth performance of weaning pigs compared to the basal diet group (p<0.05). Additionally, 0.2% MR-1 induced an elevation in the levels of mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) and showed a similar pattern ($TNF{\alpha}$ and $IFN{\gamma}$ production) to the antibiotic treated pigs. Taken together, we suggest that 0.2% MR-1 makes probiotics an alternative to antibiotics in weaning pigs.

• Journal of Dental Rehabilitation and Applied Science
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• v.25 no.2
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• pp.109-123
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• 2009

The aim of the study was to assess the influence of insertion torque of bone quality and to compare axial force, moment and von Mises stress using finite element analysis of plastoelastic property for bone stress and strain by dividing bone quality to its thickness of cortical bone, density of trabecular bone and existence of lower cortical bone when implant inserted to mandibular premolar region. The $Br{\aa}nemark$ MKIII. RP implant and cylindrical bone finite model were designed as cortical bone at upper border and trabecular bone below the cortical bone. 7 models were made according to thickness of cortical bone, density of trabecular bone and bicortical anchorage and von Mises stress, axial force and moment were compared by running time. Dividing the insertion time, it seemed 300msec that inferior border of implant flange impinged the upper border of bone, 550msec that implant flange placed in middle of upper border and 800msec that superior border of implant flange was at the same level as bone surface. The maximum axial force peak was at about 500msec, and maximum moment peak was at about 800msec. The correlation of von Mises stress distribution was seen at both peak level. The following findings were appeared by the study which compared the axial force by its each area. The axial force was measured highest when $Br{\aa}nemark$ MKIII implant flange inserts the cortical bone. And maximal moment was measured highest after axial force suddenly decreased when the flange impinged at upper border and the concentration of von Mises stress distribution was at the same site. When implant was placed, the axial force and moment was measured high as the cortical bone got thicker and the force concentrated at the cortical bone site. The influence of density in trabecular bone to axial force was less when cortical bone was 1.5 mm thick but it might be more affected when the thickness was 0.5 mm. The total axial force with bicortical anchorage, was similar when upper border thickness was the same. But at the lower border the axial force of bicortical model was higher than that of monocortical model. Within the limitation of this FEA study, the insertion torque was most affected by the thickness of cortical bone when it was placed the $Br{\aa}nemark$ MKIII implant in premolar region of mandible.

• Journal of Korean Society of Forest Science
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• v.38 no.1
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• pp.13-18
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• 1978

In this study, enzymatic hydrolysis of the holocellulose from Alnus hirsuta (Spach) Rupr. (8-14 yr's) was investigated using crude cellulase preparations of Trichoderma viride Pers. ex. Fr. SANK 16374. And conducted on the optimum condition of the treated substrate for saccharification. A strain of Trichoderma viride Pers. ex. Fr. SANK 16374 was found to be highly efficient for the cellulase productivity, especially in the submerged culture process. The culture medium used in this experiment was prepared from an extract of wheat bran consisting also of $KH_2PO_410$, $(NH_4)_2$ $SO_4$ 3, $NaNO_3$ 3, and $MgSO_4$ $7H_2O$ 0.5g/l. Cellulose powder (Toyo filter paper, 60 mesh) was found to be an importent factar for inducing the cellulase formation. And the cellulase produced in the culture fluid was salted out quantitatively by the use of ammonium sulfate (Fig. 1) Reducing sugar was determined by the Dinitrosalicylic acid (DNS) method, using reagents prepared according to the method of Sumner (1925). The results obtained were summerized as follows; 1. The method of delignification were treated by the Peracetic acid (PA) method, according to the method of Toyama (1970). The yield of holocellulose were decreased in accordance with increasing concentration of Peracetic acid solution; delignification of Alnus hirsuta Rupr. with 20% Peracetic acid was satisfied for 48 hours and 40%~60% peracetic acid was satisfied for 24 hrs: 2. The substrate (holocellulose) was changed easely into fine powder with enzymatic hydrolysis and cellulase exhibits optimum activity on the reducing sugar formation from substrate at the range of 60-100 mesh. 3. The reducing sugar formation increased in accordance with increasing dry temperature on holocellulose substrate was found to be $190{\pm}5^{\circ}C$. 4. The optimal heat treated time of holocellulose substrate was found to be 45 min. for the reducing sugar formation showed the best products. The reducing sugar formation did not show statisticaly significent diflerences at 5% levels by heat treated time for 45 min. and 60 min.

• Tuberculosis and Respiratory Diseases
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• v.50 no.1
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• pp.94-105
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• 2001

Background : Examining the biological susceptibility of Mycobacterium tuberculosis to pyrazinamide (PZA) in vitro is very difficult as PZA is inactive under normal culture conditions. The biological susceptibility test, an enzyme assay for Pzase activity, and a genetic test for pncA gene mutations, were performed in order to predict PZA resistance. Methods : 28 cultured clinical isolates of Mycobacterium tuberculosis were tested. The biological susceptibility was performed by the absolute concentration method using Lowenstein-Jensen media. The PZase activity was tested by means of Wayne's method. A 710-bp region includes the entire open reading frame of pncA was amplified and sequenced. Results : All six strains with positive PZase activity exhibited no pncA mutations with one strain showing a false resistance in the biological susceptibility test. Among the 22 strains with no PZase activity, 21 exhibited showed pncA mutations. In the biological susceptibility test, 20 strains were resistant, and one was susceptible, and the other flied to test. The mutation types varied with ten missense, one silent and one nonsense mutation 1 slipped-strand mispairing, and 6 frameshift mutations. Three strains had an adenine to guanine mutation at position -11 upstream of the start codon. Conclusion : The mutation at the pncA promotor region is frequent at -11 upstream position. Automatic sequencing of pncA is a useful tool for rapid and accurate detection of PZA resistant M. tuberculosis, and for demonstrating the epidemiological relatedness of the PZA resistant M. tuberculosis strains.

• Journal of Life Science
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• v.25 no.4
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• pp.433-440
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• 2015

Several studies suggest that regular consumption of walnuts may have beneficial effects against oxidative stress-mediated disease such as cancer. The present study reports the total phenolic and flavonoid contents, together with the antioxidant and antibacterial activities of several solvent extracts (methanol, n-hexane, ethyl acetate, n-butanol, and water) obtained from walnut (Juglans regia L.) green husk. MIC (minimal inhibitory concentration) values of the walnut extracts for 8 human pathogenic bacteria strain were determined using agar dilution method. Antioxidant activity of extracts were assessed using DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid)) assays, EC₅₀ of DPPH and ABTS scavenging activities, and determination of total phenolic and flavonoid content and its correlation with DPPH and ABTS scavenging capacities. Among the six extracts, ethyl acetate extract (EtOAc Ex) showed the highest antimicrobial activity at 3.2 mg/ml of MICs against Staphylococcus aureus SG511. Total flavonoids and polyphenol contents of EtOAc Ex were 42.48 mg of quercetin equivalents (QE)/g and 223.25 mg of gallic acid equivalents (GAE)/g respectively. The highest antioxidative potential was shown by the sample extracted with EtOAc Ex (EC₅₀=13.43 μg/ml for DPPH and EC₅₀=41.83 μg/ml for ABTS radical scavenging activity assay). These results showed that J. regia green husk extracts can be used as an easily accessible source of natural antibacterial agents and natural antioxidants.

• Korean Journal of Microbiology
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• v.28 no.3
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• pp.210-218
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• 1990

The survival and transfer of chromosomal genes coding for the synthesis of amino acids (threonine, tryptophan, histidine, leucine, methionine) and of plasmid-borne genes coding for resistance to antibiotics (chloramphenicol, kanamycin, erythromycin) by transformation in sterile and nonsterile soil (the soil was amended to 12% vol/vol with the clay mineral, montmorillonite) was studied. In pure culture, the numbers of vegetative cells of the Bacillus subtilis strains decreased by 1 to 1.5 orders of magnitude within one week, but spores of each strain showed lesser decreases. In sterile soil, the populations of vegetative cells and spores decreased by 1.5 to 3 orders of magnitude within 2 to 4 days and then showed little additional decreased. The transformation frequencies (number of transformants/numbers of donors and recipients) of individual amino acid-genes invitro ranged from $1.3{\pm}0.6{\times}10^{-6}$ to $6.0{\pm}2.36{\times}10^{-6}$, of two amino acid-genes from $8.5{\pm}0.7{\times}10^{-8}$ to $3.1{\pm}0.6{\times}10^{-7}$, and of the antibiotic-resistance genes from $1.5{\pm} 0.2{\itmes} 10^{-7}$ to $1.4{\pm} 0.4{\times} 10^{-5}$ . In sterile soil, the frequencies of transfer of individual amino acid-genes ranged from $2.0{\times} 10^{-7}$ to $2.0{\times} 10^{-5}$ and of the antibiotic-resistance genes from $2.0{\times} 10^{-7}$ to $9.4{\pm} 4.7{\times} 10^{-6}$. The transfer of two amino acid-genes in sterile soil was detected at a frequency of $2.0{\times} 10^{-6}$ to $4.5{\times} 10^{-6}$, but only in three instances. The transformation frequencies of antibiotic-resistance genes in nonsterile soil were essentially similar to those in sterile soil. However, to detect transformants in nonsterile soil, higher concentrations of antibiotics were needed, as the result of the large numbers of indigenous soil bacteria resistant to the concentration of antibiotics used in the sterile soil and in vitro studies. The results of these studies show that genes can be transferred by transformation in soil and that this mechanism of transfer must be considered in risk assessment of the release of genetically engineered microorganisms to the environment.

• The Korean Journal of Mycology
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• v.29 no.2
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• pp.79-85
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• 2001

A strain of Dictyophora echinovolvata ASI 32002 showing good fruiting body formation was selected. Analyses of chemical and nutritional components as well as antimicrobial activity of different parts of the mushroom such as mycelium, egg, and fruiting body were carried out. There were differences in the chemical compositions and the quantities depending on developmental stages of veiled lady mushroom, D. echinovolvata ASI 32002. Nitrogen, phosphate, magnesium, and calcium in inorganic chemicals were abundant in mycelium, and potassium and mineral elements were abundant in the egg and fruiting body. Mannitol and trehalose were abundant in free sugar contents. Glutamic acid and arginine in mycelium and aspartic acid and glutamic acid in egg and fruiting body were abundant in free amino acid contents. Linoleic acid, an polyunsaturated fatty acid, was abundant in all parts of the Dictyophora species, but compositions and quantities of other fatty acids varied depending on the different parts of the mushroom. It was detected that malic acid, lactic acid and acetic acid in mycelium, formic acid, acetic acid and fumaric acid in egg, and malic acid, citric acid, lactic acid, fumaric acid in fruiting body were abundant. The methanol extracts of D. echinovolvata ASI 32002 mycelium showed antifungal activity with minimal inhibition concentration (MIC) of $62{\sim}125\;{\mu}g/ml$ that was similar levels of cyclohexamide against Aspergillus awamori, Hypocrea nigricance and Trichoderma virens. The MIC of extracts from mycelium and fruiting body against Candida albicans was $250\;{\mu}g/ml$, similar to that of tetracycline. In addition to the above results, further as food additives and ingredient of cosmetics.