• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.61-68
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• 2002

The present study was examined effects of aesculetin and $O_2$concentrations on in vitro development of Hanwoo (Korean Native Cattle) embryos derived from in-vitro matured and fertilized (IVM-lVF) oocytes. The oocytes were cultured for the first 40~44 h after in vitro fertilization, then embryos of 2 to 8 cell stages were cultured under the different culture conditions for another 6 days. In experiment 1, the higher rates of morulae and blastocysts were produced in 5% $O_2$, than in 20% $O_2$(P＜0.05). There was significantly (P＜0.05) higher in embryos cultured with 1 $\mu\textrm{g}$/$m\ell$ than with 0, 5 and 10 $\mu\textrm{g}$/$m\ell$ of aesculetin. In experiment 2, the proportions of embryo developed with blastocysts and morulae plus blastocysts in 5% $O_2$, again was significantly (P＜0.05) higher in 20% $O_2$, during the culture with aesculetin and/or taurine. In the 5 and 20% $O_2$atmosphere, the inclusion of 1 $\mu\textrm{g}$/$m\ell$ aesculetin or 2.5mM taurine increased significantly (P＜0.05) the percentages of blastocysts and morulae plus blastocysts. In experiment 3, in medium with aesculetin plus PDGF and taurine plus EGF than other treatment groups, significantly (P＜0.05) higher developmental rates were obtained. Number of blastomere in balstocyst stage were also higher in medium with that than without aesculetin. However, there were no significant differences in all culture conditions. In experiment 4, the proportions of embryo developed to the morulae and blastocyst stages were significantly (P＜0.05) higher rates in medium with natural and commercial aesculetin than in control medium. No significant differences, however, were observed in between natural (71%) and commercial (70.0%) aesculetin. Number of blastomere in blastocyst stage were also higher in medium with natural and commercial aesculetin than in control medium. However, there was no effect on the number of blastomeres by these treatment. These data indicate that preimplatation embryos are very sensitive to condition that can cause oxygen concentration and show that efficiency role of aesculetin for improving bovine embryo development in vitro.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.17 no.4
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• pp.34-42
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• 2016

In order to prepare for the type approval test for the United States Coast Guard (USCG) Phase-II of Ballast Water Treatment System (BWTS), a phytoplankton mass culture was conducted in a mesocosm enclosure. We evaluated the response of the phytoplankton community after nutrient addition (+N, +P, and +NP) and investigated the development of the species with increasing culture time. After nutrient dosing, the phytoplankton population significantly (p < 0.05) increased from day 1 to day 3, depending on the nutrient treatments In particular, the specific growth rate of the phytoplankton community in the case of +NP treatment and + N treatment were estimated to be $2.47d^{-1}$ and $1.98d^{-1}$, respectively. The phytoplankton population density in the case of + NP treatment was approximately 50 times higher than that of the control group, suggesting that these treatments could be useful for mass culturing phytoplankton (> 75% of natural community) for the approval regulation of USCG Phase-II. In the phytoplankton community of the mesocosm, Pseudo-nitzchia spp. dominated in the logarithmic growth phase. The cell density decreased significantly (p < 0.05) with increasing time, coinciding with the nutrient limitation. At that time, the dominance of Pseudo-nitzchia spp. shifted to that of Cylindrotheca closterium. Therefore, the optimum nutrient concentration ($N:30{\mu}M$, $P:3{\mu}M$) and reasonable harvesting time (after 3 days in summer) found in this study for the mass culturing of phytoplankton may be helpful to meet the USCG Phase-II biological criteria to be used in BWTS.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.471-477
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• 2004

Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.

• Parasites, Hosts and Diseases
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• v.21 no.2
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• pp.234-240
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• 1983

The pathogenicity of free-living amoeba, Waegleria fcwleri, is influenced according to the strain, cultural condition and host (Culbertson et at., 1968; Carter, 1970; Wong et at., 1975), Phillips (1973) demonstrated that Entamoeba histolytica became avirulent after more than 2 year maintenance in axonic culture in vitro. This study was carried out to compare the difference in pathogenicity between two strains of N. fowleri, one of a prolonged maintenance in arsenic medium and the other one obtained by serial brain passage in mice. The 0 strain was that N. fowleri had cultivated axenically more than 7 years in CGVS medium. The 2-1 strain was obtained from the brain of mouse inoculated intranasally with a strain, which was from the mouse brain infected with 0 strain, and cultured for 15 weeks until the beginning of this experiment. White male mice weighing 18-22 g were used. Mice were anesthetized by an intraperitoneal injection of about 1 mg secobarbital, and inoculated intranasally with $10{\times}$10^4 live N. fowleri trophoBoites in a $5{\;}{\mu}l$ cell suspension. Sluggish behaviour, nervousness, rotation and leg paralysis were developed earlier and more frequently in the 2-1 experimental group than the control 0 group. Pathological changes such as inflammatory and necrotic lesion were observed in the olfactory and anterior portion of brain, and these changes were more extensive in the 2-1 group. The edematous and inflammatory changes in lung were demonstrated in mice died after 13th day post-inoculation. The experimental mice of 2-1 group began to die suddenly from 7th day post-inoculation, and the survival time in 2-1 group mice was shorter than 0 group mice. The typical primary amoebic meningoencephalitis was developed in the mice inoculated intranasally with N. fowleri. The prolonged maintenance of N. fowleri amoebae in axonic CGVS medium was observed to have lost their original pathogenicity for mice, but their pathogenicity was restored by serial brain passage in mice.

• Korean Journal of Microbiology
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• v.33 no.1
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• pp.15-21
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• 1997

We studied the characteristics of extracellular autolysins from Moraxella sp. CK-1 which has been known to lyse the cyanobacterial cell walls. This bacterium excreted autolysins from the early exponential growth phase. These enzymes showed optimal action condition of 60-$70^{\circ}C$ and pH 9.0. Whereas $Na^{+}$, $K^{+}$ and $Li^{+}$ ions exhibited positive effect on the enzyme activity, $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$ and $Mn^{2+}$ ions exhibited negative effect. Especially, $Fe^{2+}$ and $Cu^{2+}$ ions almost completely suppressed the activity. Four extracellular autolysins of 30, 32, 38 and 41 kDa were detected in renaturing SOS-PAGE gel containing 0.2% heat-killed Micrococcus luteus cells as substrate. Among these 4 autolysins, 2 enzymes of 32 and 41 kDa distributed in the culture medium throughout the experimental time, but the 38 kOa enzyme diminished and 30 kOa began to appear at mid-exponential growth phase. When SOS-insoluble peptidoglycan of M. luteus was treated with the autolysins of Moraxella sp. CK-l, the concentration of free amino groups in reaction mixture increased. This indicates that the autolysins are N-acetylmuramyl-L-alanine amidase or endopeptidase.

• Microbiology and Biotechnology Letters
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• v.13 no.1
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• pp.71-77
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• 1985

A bacterium which grows on cyclohexanol as sole carbon and energy source was isolated from sludge of industrial areas in Taegu and identified as Acinetobacter calcoaceticus C-15. The growth medium for the optimal culture condition was composed of 0.2% cyclohexanol, 0.11% $NH_4Cl$, 0.05% $KH_2PO_4$, 0.2% $K_2HPO_4$, 0.02% $MgSO_4{\cdot}7H_2O$, and 0.05% yeast extracts. The optimal pH value and temperature for the growth were 7.2 and $33^{\circ}C$, respectively. Specific growth rate of A. calcoaceticus C-15 at $33^{\circ}C$ on the cyclohexanol and cyclohexanone was $0.27hr^{-1}$ and $0.15hr^{-1}$, respectively. Growth yield for cyclohexanol was 1.0. The bacteria utilized ethanol, 1-butanol, 1-pentanol, and cyclohexanol as a carbon source but not methanol, 1-hexanol, m-cresol, glycerol, and cyclohexane. The bacteria grew on benzoate, adipate, acetate, and citrate, but did not on salicylate, phthalate, p-hydroxybenzoate, and gluconate. A calcoaceticus C-15 did not utilize all kind of sugars other than xylose. Cell-free extracts contained $NAD^+$-linked cyclohexanol dehydrogenase which catalized the oxidation of cyclohexanol to cyclohexanone.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.451-455
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• 2006

An efficient pilot scale (10 ton) three-stage methane fermentation system to digest food waste has been developed in this laboratory. This system consisted of three stages: semianaerobic hydrolysis, anaerobic acidogenesis and strictly anaerobic methanogenesis. From the secondary acidogenesis reactor, a novel strain KA4 responsible for alcohol fermentation was isolated and characterized. The cell was oval and its dimension was $5.5-6.5{\times}3.5-4.5\;{\mu}m$. This strain was identified as Saccharomyces cerevisiae KA4 by 26S rDNA D1/D2 rDNA sequence. Optimal culture temperature was $30-35^{\circ}C$. Cells were tolerant to 5% (v/v) ethanol concentration, however, were inhibited significantly by higher ethanol concentration up to 7%. The strain could grow well up to 50% (w/v) initial glucose concentration in the YM liquid medium, however, optimal concentration for ethanol fermentation was 10%. It could produce ethanol in a broad initial pH range from 4 to 10, and optimal pH was 6. In this condition, the strain converted 10% glucose to 7.4% ethanol during 24 hr, and ethanol yield was estimated to be 2.87 moi EtOH/mol glucose.

• KSBB Journal
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• v.15 no.2
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• pp.113-119
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• 2000

In order to produce xylitol with high yield, experiments were carried out to develope xylitol dehydrogenase (XDH) defective m mutant from Pichia stipitis and to investigate the xylit이 fermentation characteristics of mutant strain. After treatment of P s stipitis with EMS, mutant PXM-4 was selected based on the XDH activity and xylitol production capability. Among the tested c cosubstrates, galactose was selected as an adequate cosubstrate on xyl뻐I production of mutant PXM-4. With the increase of galactose concentration, xylitol production was decreased because the transport of xylose into cell was inhibited by g galactose. The optimal concentration of galactose for the production of xylitol using 20 g/L xylose was 20 g/L. Under this c condition, maximum concentration of xylitol and yield were 14.4 g/L and 97%, respectively. In order to prevent the inhibitory e effect of xylose transport by galactose, galactose was fed with low concentration and the concentration of xylitol produced w was increased up to 25 g/L. In the fermentation of corn cob hydrolyzate by mutant PXM-4, xylose was completely converted t to xylit이 with a 100% yield in 4 days culture.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1798-1807
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• 2002

This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Journal of the Korean Applied Science and Technology
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• v.16 no.3
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• pp.263-271
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• 1999

We checked the presence of phospholipase $A_2(PLA)_2$ which could split the ester bond at the position 2 in the glycerol backbone of glycerophospholipids, in the cells of hyperthermophiles of Pyrococcus horikoshii and Sulfolobus acidocaldarius. The results obtained are as follows; (1). Pyrococcus horikoshii cells were grown in obligate anaerobic conditions at $95^{\circ}C$ and they needed sulfur as energy source instead of oxygen, while Sulfolobus acidocaldarius species grew well in the aerobic medium (pH 2.5) containing yeast and sucrose at $75^{\circ}C$. (2). Pyrococcus horikoshii cells produced phospholipase $A_2$ in the cell culture media although this species did not show lipase activity at least in the pH range of 1.5 ${\sim}$ 3.5. Sulfolobus acidocaldarius cells produced lipase hydrolyzing triacylglycerols such as triolein, but did not split any kind of phospholipids used as substates. (3). The compound of 1-decanoyl-2-(p-nitrophenylglutaryl) phosphatidylcholine was not suitable for a substrate in this experiment, though frequently used as a subtrate for checking presence of phospholipase $A_2$, for its decomposi-tion in this experiment. The L-${\alpha}$-phosphatidylcholine-${\beta}$-[N-7-nitrobenz-2-oxa-1, 3-diazol]aminohexanoyl-${\gamma}$-hexadecanoyl labelled with a fluorescent material, did not show any migration of acyl chains in the molecule during the reaction with phospholipase $A_2$ under a hot condition. (4). Phospholipase $A_2$ in the cells of Pyrococcus horikoshii, showed the optimum activity at $pH6.7{\sim}7.2$ and $95{\sim}105^{\circ}C$, respectively, and was activated by addition of calcium chloride solution. Andthe phospholipase $A_2$ specifically hydrolyzed glycero-phospholipids such as phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine and phosphatidyl inositol, but could not split phospholipid containing ether bonds in the molecule such as DL -${\alpha}$-phosphatidylcholine-${\beta}$-palmitoyl-${\gamma}$-O-hexadecyl, DL-${\alpha}$-phosphati- dylcholine-${\beta}$- oleoyl-${\gamma}$-O-hexadecyl, DL-phosphatidylcholine-dihexadecyl.