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.
In order to obtain basic information on the production of single cell protein from petroleum, more than 400 yeast strains were isolated from various soil samples in Korea utilizing petroleum hydrocarbon as the sole carbon source. A yeast strain showing the highest cell yield among the isolated strains was selected and identified. The optimal culture condition was searched in the flasks shaken throughout the procedure. And the growing characteristics for the selected yeast strain and chemical analysis of the yeast cell component were carried out. The results obtained were as follows: 1. The selected yeast strain was identified as Candida curvata and we named it Candida curvata-SNU 70. 2. The composition of the medium proposed for the present yeast strain is: Light Gas Oil 30ml, Urea 400mg, Ammonium sulfate 100mg, Potasium phosphate (monobasic) 670mg, Sodium phosphate (dibasic) 330mg, Magnesium sulfate 500mg, Calcium carbonate 3g, Yeast extract 50mg, Tween 20 0.05ml, Tap water 1,000ml. 3. Other culture conditions employed for the yeast were pH 5.5-7.0, temp. $30^{\circ}C$ under an affluent aerobic state. 4. Addition of light gas oil in portions to the culture media as the growth proceeded was more effective, especially in the cultivation on the higher oil concentration media. 5. Studies on the propagation of the yeast cells in the light gas oil medium revealed that the yeast has the lag phase lasted 16 hours and the logarithmic growth phase covered 16 to 28 hours. The specific growth rate was about $0.22\;hr^{-1}$ and doubling time was 3.2 hrs. during the logarithmic growth phase. 6. Under the cultural condition employed, the cell yield against the amount of light gas oil (wt%) was 16.1% and the protein content of the dried yeast cells was 48.4%.
An analytical method was developed for the determination of fenpropimorph, a morpholine fungicide, in hulled rice, potato, soybean, mandarin and green pepper using QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) sample preparation and LC-MS/MS (liquid chromatography-tandem mass spectrometry). The QuEChERS extraction was performed with acetonitrile followed by addition of anhydrous magnesium sulfate and sodium chloride. After centrifugation, d-SPE (dispersive solid phase extraction) cleanup was conducted using anhydrous magnesium sulfate, primary secondary amine sorbents and graphitized carbon black. The matrix-matched calibration curves were constructed using seven concentration levels, from 0.0025 to 0.25 mg/kg, and their correlation coefficient ($R^2$) of five agricultural products were higher than 0.9899. The limits of detection (LOD) and quantification (LOQ) were 0.001 and 0.0025 mg/kg, respectively, and the limits of quantification for the analytical method were 0.01 mg/kg. Average recoveries spiked at three levels (LOQ, $LOQ{\times}10$, $LOQ{\times}50$, n=5) and were in the range of 90.9~110.5% with associated relative standard deviation values less than 5.7%. As a result of the inter-laboratory validation, the average recoveries between the two laboratories were 88.6~101.4% and the coefficient of variation was also below 15%. All optimized results were satisfied the criteria ranges requested in the Codex guidelines and Food Safety Evaluation Department guidelines. This study could serve as a reference for safety management relative to fenpropimorph residues in imported and domestic agricultural products.
In order to investigate the effects of supplemental ionic surfactants in in vitro ruminal fermentation, N-Lauroylsarcosine sodium salt(N-LSS) and sodium dodecyl sulfate(SDS) for negative(-) ionic surfactant, and hexadecylpyridinium chloride monohydrate(HPCM) and hexadecyltrimethyl ammonium bromide(HTAB) for positive (+) ionic surfactant were supplemented by 0.05% and 0.1% into the Dehority’s artificial medium containing rice straw(1mm) as a substrate. In vitro DM digestibility, the growth of rumen mixed microbes, pH, cumulative gas production and SEM(Scanning Electron Microscopy) observation of microbial attachment on rice straw particle were investigated through the experiment composing 9 treatments (two supplemental levels of two positive ionic(+) surfactant, two supplemental levels of two negative(-) ionic surfactant) including the control. The sample collection was at 6, 12, 24, 48 and 72 h post fermentation with 3 replications per treatments. DM digestibility in treatments supplemented (+) or (-) surfactants almost stopped afterward 12 h fermentation, in vitro DM digestibility at 72 h post fermentation in the ionic surfactants was at half level of that of the control(P<0.05). Accumulative gas production in in vitro was less(P<0.05) with addition of ionic surfactants compared to the control. The amount of rumen mixed microbes recovered from in vitro incubation fluid pleateaued at 12 h post fermentation for the positive (+) ionic surfactants, but steadily increased as fermentation time elapsed for the control. Rumen microbial growth rate was significantly(P<0.05) low in the negative(-) ionic surfactant compared to the control. pH of the incubation fluid was ranged from 6.02 to 7.20, and was the highest in the negative(-) ionic surfactants, and was the lowest in the control(P<0.05). In SEM observation, rumen microbial population attached on rice straw particle was less with addition of ionic surfactants than the control. In conclusion we could not found any positive effects of negative- and positive- charged surfactants on rumunal fermentation characteristics and rumen microbial growth rates.
Soil dispersion and heavy metal leaching with two heavy metal-contaminated soils were studied to derive the optimal dispersion condition in the course of developing the remedial technology using magnetic separation. The dispersion solutions of pyrophosphate, hexametaphosphate, orthophosphate and sodium dodecylsulfate (SDS) at 1 - 200 mM and the pH of solutions was adjusted to be 9 - 12 with NaOH. The clay content of suspension as an indicator of dispersion rate and the heavy metal concentration of the solution were tested at the different pHs and concentrations of the dispersion solution during the experiment. The dispersion rate increased with increasing the pH and dispersion agent concentration of the solution. The dispersion efficiency of the agents showed as follows: pyrophosphate > hexametaphosphate > SDS > orthophosphate. Arsenic leaching was sharply increased at 50 mM of phosphates and 100 mM of SDS. The adsorption of $OH^-$, phosphates and dodecysulfate on the surface of Fe- and Mn-oxides and soil organic matter and the broken edge of clay mineral might decrease the surface charge and might increase the repulsion force among soil particles. The competition between arsenic and $OH^-$, phosphates and dodecylsulfate for the adsorption site of soil particles might induce the arsenic leaching. The dispersion and heavy metal leaching data indicate that pH 11 and 10 mM pyrophosphate is the optimum dispersion solution for maximizing dispersion and minimizing heavy metal leaching.
Kim, Seung Yeol;Chung, Hai Jung;Kim, Seung Kyeom;Shin, Cheol Seung
Korean Journal of Agricultural Science
/
v.16
no.2
/
pp.225-238
/
1989
These studies were conducted to investigate the extraction, purification and characterization of oriental pear (Niitaka. Pyrus pyrifolia Nak.) protease, and the results obtained were as follows: 1. Oriental pear protease was effectively extracted by the method of homogenizing pear pulp with 0.7 volume of 0.1M-sodium phosphate buffer, pH 6.5 containing 5mM-cysteine, 40mM-2-mercaptoethanol and 2mM-EDTA at 10,000 rpm for 5 min. 2. The protease was purified by ammonium sulfate fractionation, Sephadex G-100 filtration and DEAE-Sephadex A-50 column chromatography, and the purified enzyme gave a single protein band on polyacrylamide gel electrophoresis. 3. The specific activity of purified enzyme was 29.65 unit/mg protein and the yield was 7.22%. 4. The moecular weight of the protease was estimated to be about 51,000 by SDS-polyacrylamide gel electrophoresis, and the enzyme had Km value of 54.5 mg/ml for casein. 5. The purified enzyme had a maximum activity at pH 6.0 and $50^{\circ}C$, and was stable from pH 5.5-6.5 and at temperatures below $50^{\circ}C$ 6. Casein was a better substrate for this protease compared to hemoglobin. 7. The enzyme activity was markedly inhibited by p-chloromercuribenzoic acid and heavy metal salts such as $HgCl_2$ and $MnSO_4$ also considerably inhibited the enzyme activity.
Biodegradable edible films were prepared from rice protein concentrates (RPC) made from rice wine meal by alkaline extraction and isoelectric precipitation. The effect of film forming solution pH and plasticizers were studied, and cross-linkers were added to improve mechanical properties and water vapor permeabilities (WVP) of films. Films could be formed within pH $8{\sim}11$ with tensile strength (TS) of 4.3{\sim}5.7\;MPa$. Films produced under pH 11 had the highest TS (5.7 MPa) and the lowest WVP $(0.44\;ng{\cdot}m/m^2{\cdot}s{\cdot}Pa)$. Added glycerol, polyethylene glycol 200 (PEG) and its mixture (GLY:PEG=50:50) as plasticizers also affected the mechanical properties and WVP of films. TS and elongation at break (E) of films at various plasticizer levels were $5.5{\sim}1.0\;MPa$ and $3.6{\sim}24.3%$, respectively. At the same plasticizer concentration, the highest TS was observed when glycerol was used whereas the highest E was measured when mixture was used as plasticizer. WVPs of films with thickness of $60\;{\mu}m$ were $0.39{\sim}0.54\;ng{\cdot}m/m^2{\cdot}s{\cdot}Pa$. WVP of films decreased as the ratio of glycerol/PEG 200 was decreased, and WVP increased as the total amount of plasticizer added to the films increased. Film strength was improved by the addition of small amount of sodium hydrogen sulfate, succinic anhydride, ascorbic acid and citric acid, whereas TS of films containing $0.5{\sim}2.0%$ of NaCl and $CaCl_2$ were lower than those without the salts. The highest TS (6.3 MPa) was achieved with films containing 0.1% of succinic anhydride.
A study on the application of impedance phase angle for redox titration, acid-base titration, chelate titration and precipitation titration has been carried out. A constant alternating current was passed between two platinum electrodes. One of them was a polarizable micro-electrode of $0.1cm^2$ or $0.026cm^2$ surface area and the other a non-polarizable large electrode of $1cm^2$ surface area dipped in the solution to be titrated. The impedance and the phase angle of the titration cell were measured with lock-in amplifier to obtain well behaved titration curve respectively. In titration of oxalic acid vs. potassium permanganate, the end-point was obtained successfully from the phase angle titration curve. In this experiment, the concentration of 0.0005 M to 0.05 M, the current of $50{\mu}A$ and the frequency of near 50 Hz were used. In titration of phosphoric acid vs. sodium hydroxide, the first end-point was obtained successfully on the optimum experimental condition of 0.001 M concentration, $50{\mu}A$ current and 25~97 Hz frequency. However, the end-point in titration of cupric sulfate vs. disodium-EDTA couldn't be obtained clearly. The end-point was obtained with the out-of-phase impedance curve on the experimental condition of 0.01 M concentration, $100{\mu}A$ current, 5~35 Hz frequency range. In titration of sodium chloride vs. silver nitrate, the end-point was obtained successfully on the experimental condition of 0.1 M concentration, $100{\mu}A$ current and 5~47 Hz frequency range. This study showed that the impedance phase angle was applicable for the detection of the end-points in redox titration curve, acid-base titration curve, chelate titration curve and precipitation titration curve.
From the Jungwon and Munkyeong areas which are among the famous producers of the carbonate-type groundwaters in Korea, various kinds of natural waters (deep groundwater, shallow groundwater and surface water) were collected between 1996 and 1997 and were studied for hydrogeochemical and environmental isotope (${\delta}^{34}S_{so4}$, ${\delta}^{18}O$, ${\delta}D$)systematics. Two types of deep groundwaters (carbonate type and alkali type) occur together in the two areas, and each shows distinct hydrogeochemical and environmental isotope characteristics. The carbonate type waters show the hydrochemical feature of the 'calcium(-sodium)-bicarbonate(-sulfate) type', whereas the alkali type water of the 'sodium-bicarbonate type'. The former type waters are characterized by lower pH, higher Eh, and higher amounts of dissolved ions (especialJy, $Ca^{2+}$, $Na^{+}$, $Mg^{2+}$, $HCO_3{^-}$ and $SO_4{^{2-}}$). Two types of deep groundwaters are all saturated or supersaturated with respect to calcite. Two types of deep groundwaters were both derived from pre-thermonuclear (about more than 40 years old) meteoric waters (with lighter 0 and H isotope data than younger waters, i.e., shallow cold groundwaters and surface waters) which evolved through prolonged water-rock interaction. Based on the geologic setting, water chemistry, and environmental isotope data, however, each of these two different types of deep groundwaters represents distinct hydrologic and hydrogeochemical evolution at depths. The carbonate type groundwaters were formed through mixing with acidic waters that were derived from dissolution of pyrites in hydrothermal vein ores (for the Jungwon area water) or in anthracite coal beds (for the Munkyeong area water). If the deeply percolating meteoric waters did not meet pyrites during the circulation, only the alkali type groundwaters would form. This hydrologic and hydrogeochemical model may be successfully applied to the other carbonate type groundwaters in Korea.
This study was carried out to investigate the effect of new foot-bath facility and detergent solution (sodium molylbdenate, citrate, potassium nitrate, tataric acid, sodium hypo-cholorite, and zinc sulfate) on claw health in lactating dairy cows. Minimal inhibition concentration (MIC) and minimal bactericidal concentration (MBC) of copper sulphate were 0.31% for E. coli and Bacillus isolated from cows claw. The MIC and MBC of new detergent for E. coli were 1.25% and 5%, respectively, however their respectively values for Bacillus were noticed 0.63% and 2.5%. Both 5E. coli and Bacillus populations in petri-dishes were significantly reduced (more than 95%) with the application of new detergent solution (5% or 16%). Locomotion score (LS 1-5; very good to severely bad) of lactating cows were significantly improved with in 30 days with the use of new detergent solution in foot bath. The LS2 (n=16), LS3 (n=16), and LS4 (n=7) were shown 100%, 43.8%, and 14.3% recovery rate within 30 days with the use of new detergent solution. However, LS5 (n=2) were not recovered to normal claw health and locomotion score within 30 days of new detergent application. Usage of new detergent solution for 60 days in a foot bath have shown 81.3%, 71.4% and 50.0% recovery rate in cows with LS3, LS4 and LS5, respectively. Abnormal claw incidence was reduced from 18.8% to 1.5% in overall herd (n=80) with the use of new detergent solution (16%) in a foot bath for 90 days. In conclusion, usage of 16% of our detergent solution for 60 days in a foot bath can significantly improve the cow claw health and thus mitigate the negative effects of abnormal claw on productivity of cows and dairy farm income.
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