• Korean Journal of Environmental Biology
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• v.18 no.4
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• pp.403-409
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• 2000

Surface and bottom water samples were collected from 10 stations located in the coastal area of Tongyong in April, August and October 2000. Distribution of heterotrophic bacteria, coliform bacteria and fungi in the sea water samples was investigated by measuring the corresponding viable cell number according to the plate counting method. Heterotrophic bacteria in the surface water counted 3.1$\times$10$^2$- 4.0$\times$10$^3$cfu ml$\^$-1/, 2.7$\times$10$^3$- 1.2$\times$10$\^$5/cfu ml$\^$-1/ and 1.3$\times$10$^2$- 7.2$\times$10$^2$cfu ml$\^$-1/ in April, August and October, respectively. The cell number of coliform bacteria in the surface water amounted to 0-1.5$\times$10$^1$cfu ml$\^$-1/, 3.5$\times$10$^1$- 5.2$\times$10$^3$cfu ml$\^$-1/ and 0-1.8$\times$10$^2$cfu ml$\^$-1/ in April, August and October, respectively. As for fungi, the cell number in the surface water was 0-3.0$\times$10$^1$propagules ml$\^$-1/, 3.0$\times$10$^1$- 8.0$\times$10$^1$ propagules ml$\^$-1/ and 0-2.2$\times$10$^1$ propagules ml$\^$-1/ in April, August and October respectively. The surface water samples from the station 3 in August were added with feed stuffs for fish as much as 0.01 gl$\^$-1/, 0.1 gl$\^$-1/ and 1 gl$\^$-1/ and cultured at 5$\^{C}$, 15$\^{C}$, 25$\^{C}$ and 35$\^{C}$. Microbial cells were not isolated at all when the culturing temperature was 5$\^{C}$. However, the microbial cell number increased significantly in all the surface water samples containing 1 gl$\^$-1/ of the feed stuffs when cultured at 15$\^{C}$, 25$\^{C}$ and 35$\^{C}$

• The Korean Journal of Food And Nutrition
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• v.8 no.3
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• pp.192-198
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• 1995

For the production of EMC, various professes and lipases were used to hydrolyse cheese sulk. The optimal conditions of various proteases were as follows, pronase-3$0^{\circ}C$, p14 7.0, pancreatln-4$0^{\circ}C$, pH 8.0, pacific protease-3$0^{\circ}C$, pH 7.0 and protease from Asp. sp. -5$0^{\circ}C$, pH 8.0. The optimal conditions of various lipases were as follows ; pancreatic lipase-5$0^{\circ}C$, pH 8.0, palatase ML-5$0^{\circ}C$, pH 7.0 and lipase form Candida -4$0^{\circ}C$, pH U.0. After hydrolysation under optimal conditions, the amounts of free amino acid and free fatty ac14 were increased with reaction time. Hydrolysates of pacific protease and pronase were showed high amount of free amino acid(0.67mg/ml and 0.74mg/ml). Especially EMC had high amount of glutamic acid and leucine. Lipase from Candida cylindracea produced high amount of free fatty acid (24.63 mg/ml) Butyric acrid, palmitic acid, stearic acid and oleic acid among free fatty acids were showed high amounts. Sensory evaluation of various MC were tasted nth 8 panelist. EMC produced with pancreatic lipase was most bitterness and EMC produced with palatase ML was best acceptable cheese flavor.

• Journal of Life Science
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• v.24 no.4
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• pp.419-427
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• 2014

In order to examine antioxidative characteristics of Artemisia capillaris response surface methodology was used to optimize the hot water extraction process by analyzing and monitoring the extraction condition. For total phenolic compounds content, the optimal extraction temperature, time and amount of solvent per sample were $94.50^{\circ}C$, 2.06 hr and 25.03 ml/g, respectively. Also, the optimal conditions for electronic donating ability were $91.82^{\circ}C$, 2.90 hr and 20.88 ml/g, respectively. The nitrile scavenging ability (pH 1.2) was optimized using the extraction temperature of $97.36^{\circ}C$, extraction time 2.75 hr and 15.19 ml/g as the amount of solvent per sample. Regression equations of total phenolic compounds content, electron donating ability and nitrile scavenging ability as dependent variable were deduced from each analyzed extraction condition. And finally, their response surfaces were superimposed with the optimal conditions to obtain values for each extraction process factor. The predicted results through superimposing were extraction temperature $90{\sim}95^{\circ}C$, extraction time 2.5~3.5 hr and amount of solvent per sample 17~24 ml/g.

• Journal of Preventive Medicine and Public Health
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• v.28 no.4 s.51
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• pp.863-873
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• 1995

This report describes a preperation and characterization of canine blood lead(Pb) standard reference material(SRM). Three adult beagle dogs(A, B, and C)were orally dosed with gelatin capsules containing $Pb(NO_3)_2$, equivalent to $10\sim80mg$ Pb/kg body weight. Blood was drawn 24 hours after the dose from the cephalic vein into lead free 500ml Pyrex beaker in which EDTA.K was contained as an anticoagulant. The amount of lead given to individual dog was varied arbitrarily. Three month later, 3 canine animals were orally dosed with lead secondarily to make mixed SRM(D1) which was mixed different concentrations of lead in bloods with A1, B1, and C1 in vitro. The SRMs for A, B, C, A1, B1, C1, and D1 were distributed 2ml each into more than 300 lead free bottles, and were stored in refregerator at $4^{\circ}C$. The amount of lead in canine whole blood samples were determined using a Varian 30A atomic absorption spectrophotometer(AAS) with a model GTA-96 graphite tube atomizer with D2 background correction and a Hitachi Z-8100 AAS with Zeeman background correction. The sensitivity and detection limits for lead determination of Varian 30A were $0.46{\mu}g/L,\;0.34{\mu}g/L,\;and\;0.56{\mu}g/L,\;0.14{\mu}g/L$ of Hitachi Z-8100, respectively. Day to day variations in determination of blood lead concentration in a certain sample were $31.11{\pm}1.36{\mu}g/100ml$ by Varian 30A, and $33.08{\pm}0.82{\mu}g/100ml$ by Hitachi Z-8100, showing the difference of 3% between the two results. At the blood lead concentrations of $56.31{\pm}1.98{\mu}g/100ml(A),\;40.89{\pm}0.80{\mu}g/100ml(B),\;59.01{\pm}1.38{\mu}g/100ml(C)$, the precisions of replicated measurements by AAS were 3.52%, 1.96%, and 2.34%, respectively. Coefficient variation(CV) of SRMs(A, B, and C) within a standard sample were ranged from 0.92% to 7.50%, and those between 5 standard samples were 1.21%, 2.64%, and 1.11%, respectively, showing inter-vial variation of $1{\mu}g/100ml$. Lead levels in SRMs during one month storage were unchanged. The overall recoveries were $89.6\sim100.4%,\;91.6\sim101.9%,\;90.3\sim100.0%$ for A, B, and C SRMs, means were $56.46{\pm}2.69{\mu}g/100ml,\;39.35{\pm}1.89{\mu}g/100ml,\;57.40{\pm}2.31{\mu}g/100ml$, and measurement ranges were$52.88{\pm}59.26{\mu}g/100ml,\;37.47{\pm}41.68{\mu}g/100ml,\;54.80{\pm}60.69{\mu}g/100ml$, respectively. Those results were laid within confidence limits values. The lead concentrations in the mixed sample(D1) stored over one month period were ranged from $32.76{\mu}g/100ml\;to\;33.54{\mu}g/100ml$, with CV ranging from 1.2% to 2.7%. The results were similiar to each of single samples(A1, B1, and C1) in respect of homogeneity and stability. Results of the mixed blood sample analysed after 1 month storage at $4^{\circ}C$ by four other laboratories(L1, L2, L3, L4) were similar with those of our laboratory($L5;31.18{\pm}0.24{\mu}g/100ml$, acceptable range by $CDC;25.18\sim37.18{\mu}g/100ml$), showing the concentrations of $25.91{\pm}1.19{\mu}g/100ml(L1),\;34.16{\pm}0.22{\mu}g/100ml(L2),\;35.68{\pm}0.85{\mu}g/100ml(L3),\;30.95{\pm}0.46{\mu}g/100ml(L4)$ in a each samples.

• Korean Journal of Food Science and Technology
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• v.24 no.4
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• pp.393-395
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• 1992

The ${\alpha}-lactalbumin({\alpha}-la)$ concentration in raw and laboratory-heated milks by HPLC was 1.20 mg/ml (unheated), 1.17 mg/ml ($63^{\circ}C$, 30min), 1.13 mg/ml ($72^{\circ}C$, 15sec) and 0 mg/ml ($100^{\circ}C$, 10min), respectively. Whereas, ${\alpha}-lactalbumin$ concentration ranges of commercial milks were $1.00{\sim}1.02\;mg/ml$ (pasteurized), $0.23{\sim}0.68\;mg/ml$ (UHT-pasteurized) and $0.77{\sim}0.89\;mg/ml$ (UHT-sterilized), respectively. It was supposed that the ${\alpha}-lactalbumin$ content of sterilized milk will be lower than that of UHT milk, but the opposite occurred. This discrepancy would be caused by the different heating system in the milk plants, where indirect UHT-treatment had more heat intensity than direct UHT-processing. The HPLC determination of ${\alpha}-lactalbumin$ may be an indicator to evaluate correctly and rapidly heated milks.

• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.215-223
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• 2003

This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

• Journal of environmental and Sanitary engineering
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• v.16 no.4
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• pp.62-68
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• 2001

The advantages of hydrogen peroxide dissolution method were no discharge of noxious matter when dissolution of iron wire which used as the center supporter, reactions occur in room temperature and easy to recover dissolved iron. This study was aimed at gathering the basic data of iron wire dissolution- recovery process and proposes the reaction condition of iron wire dissolution- recovery process rind the factors influencing those reactions. The results were as follows : 1 . Hydrogen peroxide dissolution method used hydrochloric acid as the catalyst. 1. In the dissolution of iron wire(1.668 g), the condition of reaction was E1702(30 ml), HCI(20 ml) and $H_2O$(200 ml) ; time of the reaction was 18 min. P.W.(Piece weight) was 7.75 mg, and C.R. was $2.34{\;}{\Omega}$ 2. In the dissolution of iron wire(1.529 g), the condition of reaction was H7O2(30 ml), HCI(20 ml) and $H_2O$(200 ml), time of the reaction was 21 min., P.W.(Piece weight) was 7.73 mg, and C.R. was $2.35{\;}{\Omega}$. Hydrogen peroxide dissolution method used sulfuric acid as the catalyst. 1. In the dissolution of iron wire(0.834 g), the condition of reaction was $H_2O$(65 ml), $H_2SO_4$(5 ml) and 1702(5 ml) ; time of the reaction was 5 min.30 sec, P.W.(Piece weight) was 7.74 mg, and C.R. was $2.33{\;}{\Omega}$ 2. In the dissolution of iron wire(1.112 g), the condition of reaction was $H_2O$(65 ml), $H_2SO_4$(5 ml) and $H_2O_2$(5 ml) ; time of the reaction was 4 min.30 sec, P.W.(Piece weight) was 7.75 mg, and C.R. was $2.33{\;}{\Omega}$. Hydrogen peroxide dissolution method used hydrochloric acid and sulfuric acid as the catalyst confirmed a clean technology, because there were not occurred a pollutant discharged in the existing method.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.63-63
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• 2003

This study was carried out to investigate the effects of liquid boar sperm stored at 4$^{\circ}C$ on sperm motility, normal acrosome, and in-vitro fertilization and culture of pig oocytes matured in-vitro. The sperm-rich fraction (30~60 ml) of ejaculate was collected into an insulated vacuum bottle. Semen was slowly cooled to room temperature (20~23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min at 800$\times$g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of lactose, egg yolk and N-acetyl-D-glucosamine (LEN) diluent to provide 1.0$\times$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$ and preserved for 5 days to examine sperm motility and normal acrosome. The medium used for oocyte maturation was modified tissue culture medium (TCM) 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 6 h in 500 ${mu}ell$ mTBM fertilization media with 0.2, 1, 5 and 10$\times$10$^{6}$ /ml sperm concentration, respectively. At 6 h after IVF, oocytes were transferred into 500 ${mu}ell$ Hepes-buffered NCSU-23 culture medium for further culture of 6, 48 and 144 h. There were significant differences in sperm motility and normal acrosome among preservation days and incubation times, respectively. The rates of sperm penetration and polyspermy were higher in 5 and 10$\times$10$^{6}$ sperm/ml than in 0.2 and 1$\times$10$^{6}$ sperm/ml. Male pronuclear formation was lower in 0.2$\times$10$^{6}$ sperm/ml than in 1, 5 and 10$\times$10$^{6}$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in 10$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. The rate of blastocysts from the cleaved oocytes (2~4 cell stage) was highest in 1$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend 1$\times$10$^{6}$ ml sperm concentration for in-vitro fertilization of pig oocytes.

• Journal of Marine Life Science
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• v.7 no.2
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• pp.94-101
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• 2022

In this study, the marine medaka Oryzias javanicus was exposed to two concentrations of non-toxin-producing red tide dinoflagellate C. polykrikoides (1,000 and 2,000 cells ml^-1) for 96 h, and the time-course biochemical responses of antioxidant and immunity parameters were analyzed in the liver tissue. Significant ichthyotoxicity with increasing cell concentrations of C. polykrikoides and exposure period was observed for 96 h. Opercular respiratory rate was lowered in marine medaka exposed to 2,000 cells ml^-1 of C. polykrikoides. Intracellular malondialdehyde (MDA) content significantly elevated in response to both cell concentrations. In the case of glutathione (GSH) content, the levels were significantly elevated by 1,000 cells ml^-1 of C. polykrikoides, but the contents significantly depleted upon exposure to 2,000 cells ml^-1 of C. polykrikoides. Similarly, enzymatic activities of catalase (CAT) and superoxide dismutase (SOD) were increased by 1,000 cells ml^-1 of C. polykrikoides, whereas their activities were lowered by 2,000 cells ml^-1 of C. polykrikoides. Analysis of the two immunity parameters, alternative complement pathway and lysozyme, showed significantly lowered activities in 2,000 cells ml^-1 of C. polykrikoides-exposed liver tissue. These biochemical effects of C. polykrikoides on marine medaka would be helpful for understanding its acute effects in marine fish.

• The Korean Journal of Zoology
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• v.29 no.3
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• pp.181-189
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• 1986

The present study was undertaken to investigate whether the known adenylate cyclase activators, forskolin and cholera toxin, would affect the germinal vesicle breakdown (GVBD) and the production of cAMP in mouse oocytes in vitro. To do this, in vitro oocyte culture method and adenylate cyclase assay were employed. In response to different concentrations of forskolin (20 to 80 $\\mu$g/ml) added to a culture medium, the percentage of GVBD significantly decreased (56 to 31%) in a dose-dependent manner as compared to that of control (63%). This inhibitory phenomenon by forskolin was reversible since the rate of GVBD was returned to the control level when the oocytes were transferred to a control medium following exposure to forskolin (80 $\\mu$g/ml). Treatment of cholera toxin (10 to 1, 000 ng/ml) was, however, ineffective in suppressing GVBD. When forskolin (10 to 80 $\\mu$g/ml) was added to the mouse oocyte extracts, cAMP production significantly increased by 5 to 18 fold, whereas cholera toxin (10 to 1, 000 ng/ml) was no longer effective. In addition, treatment of guanidyl-imidodiphosphate (GppNHp, 100 $\\mu$M), which is an activator of the regulatory unit of adenylate cycleas, with forskolin did not exhibit any changes in cAMP production as compared to that induced by forskolin alone. Neither cholera toxin nor cholera toxin plus GppNHp (100 $\\mu$M) exhibited any differences in mouse oocytes. From the above results, the suppression of GVBD by forskolin may be mediated by a high level of intracellular cAMP in mouse oocytes. It appears that the changes in intracellular cAMP level may an important role in the mouse oocyte maturation.