• Applied Biological Chemistry
• /
• 제11권
• /
• pp.1-27
• /
• 1969

More than thirty kinds of sea food pickles have been eaten in Korea. Out of these salted yellow tail pickle, salted clam pickle, salted oyster pickle, and salted cuttlefish pickle were employed for the analysis of their components, identification of main fermenting microbes, and determination of enzyme characteristics concerned. Also studied was the effect of enzymic action of microbes, which are concerned with the fermenting of pickles, on the production of flavorous 5'-mononucleotides and amino acids. The results are summarized as follows: 1. Microflora observed in the pickles are: (a) Total count of viable cells after 1-2 months of pickling was found to be $10^7$ and that after 6 months decreased to $10^4$. (b) Microbial occurence in the early stage of pickling was observed to be 10-20% Micrococcus spp., 10-20% Brevibacterium spp., 0-30% Sarcina spp., 20-30% Leuconostoc spp., ca 30% Bacillus spp., 0-10% Pseudomonas spp., 0-10% Flavobacterium spp., and 0-20% yeast. (c) Following the early stage of pickling, mainly halophilic bacteria such as Bacillus subtilis, Leuconostoc mesenteroides, Pediococcus halophilus and Sarcina litoralis, were found to exhibit an effect on the fermentation of pickle and their enzyme activities were in direct concern in fermentation of pickles. (d) Among the bacteria participating in the fermentation, Sarcina litoralis 8-14 and 8-16 strains were in need of high nutritional requirement and the former was grown only in the presence of purine, pyrimidine and cystine and the latter purine, pyrimidine and glutamic acid. 2. Enzyme characteristics studied in relation to the raw materials and the concerned microbes isolated are as follows: (a) A small amount of protease was found in the raw materials and 30-60% decrease in protease activity was demonstrated at 7% salt concentration. (b) Protease activity of halophilic bacteria, Bacillus subtilis 7-6, 11-1, 3-6 and 9-4 strains, in the complete media decreased by 10-30% at the 7% salt concentration and that of Sarcina litoralis 8-14 and 8-16 strains decreased by 10-20%. (c) Proteins in the raw materials were found to be hydrolyzed to yield free amino acids by protease in the fermenting microbes. (d) No accumulation of flavorous 5'-mononucleotides was demonstrated because RNA-depolymerase in the raw materials and the pickles tended to decompose RNA into nucleoside and phosphoric acid. (e) The enzyme produced in Bacillus subtilis 3-6 strain isolated from the salted clam pickles, was ascertained to be 5'-phosphodiesterase because of its ability to decompose RNA and thus accumulating 5'-mononucleotide. (f) It was demonstrated that the activity of phosphodiesterase in Bacillus subtilis 3-6 strain was enhanced by some components in the corn steep liquor and salted clam pickle. The enzyme activity was found to decrease by 10-30% and 40-60% at the salt concentration of 10% and 20%, respectively. 3. Quantitative data for free amino acids in the pickles are as follows: (a) Amounts of acidic amino acids such as glutamic and aspartic acids in salted clam pickle, were observed to be 2-10 times other pickles and it is considered that the abundance in these amino acids may contribute significantly to the specific flavor of this food. (b) Large amounts of basic amino acids such as arginine and histidine were found to occur in salted yellow tail pickle. (c) It is much interesting that in the salted cuttlefish pickle the contents of sulfur-containing amino acids were exceedingly high compared with those of others: cystine was found to be 17-130 times and methionine, 7-19 times. (d) In the salted oyster pickle a high content of some essential amino acids such as lysine, threonine, isoleucine and leucine, was demonstrated and a specific flavor of the pickle was ascribed to the sweet amino acids. Contents of alanine and glycine in the salted oyster pickle were 4 and 3-14 times as much as those of the others respectively. 4. Analytical data for 5'-mononucleotides in the pickles are as follows: (a) 5'-Adenylic acid and 3'-adenylic acid were found in large amounts in the salted yellow tail pickle and 5'-inosinic acid in lesser amount. (b) 5'-Adenylic acid, especially 3'-adenylic acid predominated in amount in the salted oyster pickle over that in the other pickles. (c) The salted cuttlefish pickle was found to contain only 5'-adenylic acid and 3'-adenylic acid. It has become evident from the above fact that clam and the invertebrate lack of adenylic deaminase and contain high content of adenylic acid. Thus, they were demonstrated to be the AMP-type. (d) 5'-Inosinic acid was contained in the salted yellow tail pickle in a significant concentration, and it might be considered to be IMP-type. 5. Comparative data for flavor with regard to the flavorous amino acids and the contents of 5'-mononucleotides are: (a) A specific flavor of salted yellow tail pickle was ascribed to the abundance in glutamic acid and aspartic acid, and to the existence of a small amount of flavorous 5'-inosinic acid. The combined effect of these components was belived to exhibit a synergistic action in producing a specific fiavor to the pickle. (b) A specific flavor of salted clam pickle has been demonstrated to be attributable to the richness in glutamic acid and aspartic acid rather than to that of 5'-mononucleotides.

• Applied Biological Chemistry
• /
• 제21권3호
• /
• pp.150-184
• /
• 1978

Nutritional characteristics and physio-chemical properties of mycelial growth and fruitbody formation of oyster mushroom(Pleurotus ostreatus)in synthetic media, the curtural condition for the commerical production in the rice straw and poplar sawdust media, and the changes of the chemical components of the media and mushroom during the cultivation were investigated. The results can be summarized as follows: 1. Among the carbon sources mannitol and sucrose gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while lactose and rhamnose gave no mycelial growth. Also, citric acid, succinic acid, ethyl alcohol and glycerol gave poor fruit-body formation, and acetic acid, formic acid, fumaric acid, n-butyl alcohol, n-propyl alcohol and iso-butyl alcohol inhibited mycelial growth. 2. Among the nitrogen sources peptone gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while D,L-alanine, asparatic acid, glycine and serine gave very poor fruit-body formation, and nitrite nitrogens, L-tryptophan and L-tyrosine inhibited mycelial growth. Inorganic nitrogens and amino acids added to peptone were effective for fruit-body growth, and thus addition of ammonium sulfate, ammonium tartarate, D,L-alanine and L-leucine resulted in about 10% increase fruit-body yield. L-asparic acid about 15%, L-arginine about 20%, L-glutamic acid, and L-lysine about 25%. 3. At C/N ratio of 15.23 fruit-body formation was fast, but the yield decreased, and at C/N ratio of 11.42 fruit-body formation was slow, but the yield increased. Also, at the same C/N ratio the higher the concentration of mannitol and petone, the higher yield was produced. Thus, from the view point of both yield of fruit-body and time required for fruiting the optimum C/N ratio would be 30. 46. 4. Thiamine, potassium dihydrogen phosphate and magnecium sulfate at the concentration of $50{\mu}g%$. 0.2% and 0.02-0.03%, respectively, gave excellent mycelial and fruit-body growth. Among the micronutrients ferrous sulfate, zinc sulfate and manganese sulfate showed synergetic growth promoting effect but lack of manganese resulted in a little reduction in mycelial and fruit-body growth. The optimum concentrati on of each these nutrients was 0.02mg%. 5. Cytosine and indole acetic acid at 0.2-1mg% and 0.01mg%, respectively, increased amount of mycelia, but had no effect on yield of fruit-body. The other purine and pyrimidine bases and plant hormones also had no effect on mycelial and fruit-belly yield. 6. Illumination inhibited mycelial growth, but illumination during the latter part of vegetative growth induced primordia formation. The optimum light intensity and exposure time was 100 to 500 lux and 6-12 hours per day, respectively. Higher intensity of light was injurous, and in darkness only vegetative growth without primordia formation was continued. 7. The optimum temperature for mycelial growth was $25^{\circ}C$ and for fruit-body formation 10 to $15^{\circi}C$. The optimum pH range was from 5.0 to 6.5. The most excellent fry it-body formation were produced from the mycelium grown for 7 to 10 days. The lesser the volume of media, the more rapid the formation of fruit-body; and the lower the yield of fruit-body; and the more the volume of media, the slower the formation of fruit-body, and the higher the yield of fruit-body. The primordia formation was inhibited by $CO_2$. 8. The optimum moisture content for mycelial growth was over 70% in the bottle media of rice straw and poplar sawdust. 10% addition of rice bran to the media exhibited excellent mycelial growth and fruit-body formation, and the addition of calciumcarbonate alone was effective, but the addition of calcium carbonate was ineffective in the presence of rice bran. 9. In the cultivation experiments the total yield of mushroom from the rice straw media was $14.99kg/m^2$, and from the sawdust media $6.52kg/m^2$, 90% of which was produced from the first and second cropping period. The total yield from the rice straw media was about 2.3 times as high as that from the sawdust media. 10. Among the chemical components of the media little change was observed in the content of ash on the dry weight basis, and organic matter content decreased as the cultivation progressed. Moisture content, which was about 79% at the time of spawning, decreased a little during the period of mycelial propagation, after which no change was observed. 11. During the period from spawning to the fourth cropping about 16.7% of the dry matter, about 19.3% of organic matter, and about 40% of nitrogen were lost from the rice straw media; about 7.5% of dry mallet, about 7.6% of organic matter, and about 20% of nitrogen were lost from the sawdust media. For the production of 1kg of mushroom about 232g of organic matter and about 7.0g of nitrogen were consumed from the rice straw media; about 235g of organic matter and about 6.8g of nitrogen were consumed from the sawdust media, 1㎏ of mushroom from either of media contains 82.4 and 82.3g of organic matter and 5.6 and 5.4g of nitrogen, respectively. 12. Total nitrogen content of the two media decreased gradually as the cultivation progressed, and total loss of insoluble nitrogen was greater than that of soluble nitrogen. Content of amino nitrogen continued to increase up to the third cropping time, after which it decreased. 13. In the rice straw media 28.0 and 13.8% of the total pentosan and ${\alpha}$-cellulose, respectively, lost during the whole cultivation period was lost during the period of mycelial growth; in the sawdust media 24.1 and 11.9% of the total pentosan and ${\alpha}$-cellulose, respectively, was lost during the period of mycelial growth. Lignin content in the media began to decrease slightly from the second cropping time, while the content of reduced sugar, trehalose and mannitol continued to increase. C/N ratio of the rice straw media decreased from 33.2 at spawining to 30.0 at ending; that of the sawdust media decreased from 61.3 to 60.0. 14. In both media phosphorus, potassium, manganese and zinc decreased, at magnesium, calcium and copper showed irregular changes, and iron had a tendency to be increased. 15. Enzyme activities are much higher in the rice straw media than in the sawdust media. CMC saccharifying and liquefying activity gradually increased from after mycelial propagation to the second cropping, after which it decreased in both media. Xylanase activity rapidly and greatly increased during the second cropping period rather than the first period. At the start of the third cropping period the activity decreased rapidly in the rice straw media, which was not observed in the sawdust media. Protease activity was highest after mycelial propagation, after which it gradually decreased. The pH of the rice straw media decreased from 6.3 at spawning to 5.0 after fourth cropping; that of the sawdust media decreased from 5.7 to 4.9. 16. The contents of all the components except crude fibre of the mushroom from the rice straw media were higher than those from the sawdust media. Little change was observed in the content of the components of mushroom cropped from the first to the third period, but slight decrease was noticed at the fourth cropping.

• Journal of Chest Surgery
• /
• 제37권12호
• /
• pp.967-977
• /
• 2004

Background: DNA methylation is one of the important gene expression mechanisms of the cell. When cytosine of CpG dinucleotide in promotor is hypomethylated, expression of some genes that is controlled by this promoter is altered. In this study, the author investigated the effect of DNA demethylating agent, 5-aza-2'-deoxycytidine (ADC), on the expressions of cancer antigen genes, MHC and B7 in 4 lung cancer cell lines, NCIH1703, NCIH522, MRC-5, and A549. Material and Method: After treatment of cell lines, NCIH1703, NCIH522, MRC-5 and A549 with ADC (1 uM) for 48 hours, RT-PCR was performed by using the primers of MAGE, GAGE, NY-ESO-1, PSMA, CEA, and SCC antigen gene. In order to find the optimal ADC treatment condition for induction of cancer antigen, we studied the effect of ADC treatment time and dose on the cancer antigen gene expression. To know the effect of ADC on the expression of MHC or B7 and cell growth, cells were treated with 1 uM of ADC for 72 hours for FACS analysis or cells were treated with 0.2, 1 or 5 uM of ADC for 96 hours for cell counting. Result: After treatment of ADC (1 uM) for 48 hours, the expressions of MAGE, GAGE, NY-ESO-1, and PSMA genes increased in some cell lines. Among 6 MAGE isotypes tested, and gene expression of MAGE-1, -2, -3, -4 and -6 could be induced by ADC treatment. However, CEA gene expression did not change and SCC gene expression was decreased by ADC treatment. Gene expression was generally induced 24 - 28 hours after ADC treatment and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC ADC teatment, and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC teatment in ADC-Free medium. Most gene expression could be induced at 0.2 uM of ADC, but gene expression increased dependently on ADC treatment dose. The expression of MHC and B7 was not increased by ADC treatment in all four cell lines, and the growth rate of 4 cell lines decreased significantly with the increase of ADC concentrations. Conclusion: Treatment of lung cancer cell lines with ADC increases the gene expression MAGE, GAGE and NY-ESO-1 that are capable of induction of cytotoxic T lymphocyte response. We suggest that treatment with 1 uM of ADC for 48 hours and then culturing in ADC-free medium is optimal condition for induction of cancer antigen. However, ADC has no effect on MHC and B7 induction, additional modification for increase of expression of MHC, B7 and cytokine will be needed for production of efficient cancer cell vaccine.

• Journal of Animal Science and Technology
• /
• 제48권1호
• /
• pp.29-38
• /
• 2006

The objective of this study was to estimate genetic variances of growth curve parameters in Hanwoo cows. The data used in this study were records from 1,083 Hanwoo cows raised at Hanwoo Experiment Station, National Livestock Research Institute(NLRI). First evaluation model(Model I) fit year-season of birth and age of dam as fixed effects and second model(Model II) added age at the final weight as a linear covariate to Model I. Heritability estimates of A, b and k from Gompertz model were 0.22, 0.11 and 0.07 using modelⅠ and 0.28, 0.11 and 0.12 using modelⅡ. Those from Von Bertalanffy model were 0.22, 0.11 and 0.07 using modelⅠ, 0.28, 0.11 and 0.12 using modelⅡ. Heritability estimates of A, b and k from Logistic model were 0.14, 0.07 and 0.05 using modelⅠ, 0.18, 0.07 and 0.12 using modelⅡ. Heritability estimates of A from Gompertz model were higher than those from Von Bertalanffy model or Logistic model in both model Ⅰand model Ⅱ. Heritability estimates of b from Logistic model were higher than those from Gompertz model or Von Bertalanffy model in both modelⅠand model Ⅱ. Heritability estimates of birth weight, weaning weight, 3 month weight, 6 month weight, 9 month weight, 12 month weight, 18 month weight, 24 month weight, 36 month weight were after linear age adjustment 0.27, 0.11, 0.19, 0.14, 0.16, 0.23, 0.52 and 0.32, respectively. Heritability estimates of birth weight, weaning weight, 3 month weight, 6 month weight, 9 month weight and 24 month weight fit by Gompertz model were larger than those estimated from linearly adjusted data. Heritability estimates of 12 month weight, 18 month weight and 36 month weight fit by Von Bertalanffy model were larger than those estimated from linearly adjusted data. In the multitrait analyses for parameters from Gompertz model, genetic and phenotypic correlations between A and k parameters were －0.47 and －0.67 using modelⅠand －0.56 and －0.63 using model Ⅱ. Those between the A and b parameters were 0.69 and 0.34 using modelⅠand 0.72 and 0.37 using model Ⅱ. Those between the b and k parameters were －0.26 and 0.01 using modelⅠand －0.30 and 0.01 using model Ⅱ. In the multitrait analyses for parameters from Von Bertalanffy model, genetic and phenotypic correlations between A and k parameters were －0.49 and －0.67 suing model Ⅰ and －0.57 and －0.70 using modelⅡ. Those between the A and b parameters were 0.61 and 0.33 using modelⅠ and 0.60 and 0.30 using model Ⅱ. Those between the b and k parameters were －0.20 and 0.02 using modelⅠ and 0.16 and 0.00 using modelⅡ. In the multitrait analyses for parameters from Logistic model, genetic and phenotypic correlations between A and k parameters were －0.43 and －0.67 using model Ⅰ and －0.50 and －0.63 using modelⅡ. Those between the A and b parameters were 0.47 and 0.22 using modelⅠ and 0.38 and 0.24 using modelⅡ. Those between the b and k parameters were －0.09 and 0.02 using model Ⅰ and －0.02 and 0.13 using model Ⅱ.

• Korean Journal of Animal Reproduction
• /
• 제27권3호
• /
• pp.215-223
• /
• 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 Korean Society of Forest Science
• /
• 제45권1호
• /
• pp.26-36
• /
• 1979

Through several trials that has done for making the fertilizing-counter plan on the soil improving species, some results have been got as follows; 1. In the non-phosphorus dressing plots soil improving species have very poor survial ratio and show us to be died step by step. It may be resons that root can not make the nodule in case of non-phosphorus dressing and so tree could not absorb the nitrogen nutrient fixed by the nodule. And root competition with the weedy speces for utilizing the nutrient and oxygen in the soil could be reasons when planting in the heavy weedy rooting site. 2. Triple super phosphate, Fused Mg Phosphate and Fused super phosphate have showed the remarkable effects on the growth of soil improving species within 3rd year after planting. But Koreaan tablet fertilizer(9-12-4) for forest purpose have reacted considerably lower effect in comparision with the above powder and grain type phosphorous fertilizer. 3. In case of tablet type fertilizer tree root will have very little phosphorus absorbing surface because phosphorus can be utilized only from the tablet surface and root can not penetrate into the tablet. This my be reson to show the poor dressing reaction of tablet fertilizer but tablet fertilizer has a possibility to be utilized during long years as a sympton in photo 6. So tablet fertilizer can have a recommendation to dress much fertilizer at p]anting year and then tree root can get much more chance for absorbing the phosphorus that could keep the high survival and for utilizing it during many years without after dressing. 4. The granurar and powder type phosphate can develop the dense root mat like photo 8 because of giving the large surface for absorbing the phosphorus and weedy root can approch to the nodule for taking the nitrogen element. So this type seems to present better effect than tablet type to control the soil movement, stem weight as 200g per meter(l meter long${\times}$0.1m width). When added the lime any effect could not be found and rather give the negative effect. So Lespedeza seed sowing and phosphorus dressing system seems us to be very reasonable method for covering the raw material of basket making, fodder and fuel wood supply. 7. Fused Mg phosphate and Fused super phosphate are good fertilizer to the soil improving species and dressing more than 30g per seedling can be recommendable amount. 5. In the unproductive and dry soil with phosphorus fertilizer Robinia pseudoacacia and Alnus firnui can grow more than 2.3m in height at 3rd year and Alnus inokumae have the rapid height growth that is more than 1.8m at 2nd year. Depending on the growth situation like the above example minirotated management has possibilities and rapid covering of erosed land can be done with the soil improving species and phosphorus fertilizer. 6. In the Lespedeza sowing plot with 40g Fused Mg phosphate dressing per meter in the eroded and unproductive forest soil Lespedeza have completely covered this poor land and produced the green.

• Journal of Korean Society of Forest Science
• /
• 제13권1호
• /
• pp.25-39
• /
• 1971

Recently successful induction of haploid plant by means of anther culture method has become a big topic among geneticists and plant breeders. The haploid plant can be used as a precious material for such basic researches as mutation or genetics. Once the haploid is obtained, production of homozygous plant is not a difficult problem. The method of producing homozygous plant can, also, be applied to the practical breeding works. When applied to the hybridization of self-fertilizing breeding period would be greatly shortened and in cross-fertilizing vegetables production of uniform hybrid seed would be very easily obtained. Last few years many scientists attempted anther cultures using various plant species, but it was successful only in several species. Unlike the other tissue cultures which use somatic organs or tissues as explants, anther culture seems to be very difficult because the plants or calli have to be induced from the haploid microspores or pollen grains. In the present experiment anther culture of fruit trees and ornamental shrubs of four genera and seven species was attemped. Anthers of Various stages ranging from tetrad and late microspore were cultured on the modified Murashige and Skoog's medium supplemented with various concentrations of auxins and kinetin as growth regulators. Handling of materials, sterilization, and other operations of culture were done by routine methods. The results were summarized as follows: 1. Calli were induced in the anthers of Forsythia Koreana Nak., Rhododendron mucronuratum Turcz., R. yedoense Max. var. Poukhanense Nak., and Prunus armeniaca L. var. ansu Max. No signs of callus were observed in Prunus persica Sieb. et Zucc. var. vurgaris Max., Pyrus ussuriensis var. macrostipes (Nak.), and Prunus salcina Lindley. 2. Calli were easily formed in any of the media with differing concentrations of auxins and kinetin. 3. In F. Koreana calli developed from anther surface and connective. Callus emerging out of anther locule was not observed. 4. Somatic calli arose from filament, connective, and inside of anther wall in R. mucronulatum. Many of the microspores accumulated starch grains. 5. The anther lobes located opposite the filament of R. yedoense turned easily to calli. This phenomenon was not observed in R. mucronulatum. Microspore embedded for a period in the medium became starch pollen. No callus was observed arising from microspore. 6. In P. armeniaca calli were not induced from somatic anther tissues. Instead, callus emerged out of anther locule rupturing the anther slit. Starch was not formed in the microspore. 7. In P. persica, Pyrus ussuriensis, and P. salcina, calli were not observed in the anthers examined more than 60 days after culture. Microspores of these species, however, were free of starch grains even after long period of subculture. 8. It was learned that somatic calli of the species examined arose usually from endothelium of anther wall, septum of two neighboring anther locules, parenchyma tissues of connectives, or anther lobes. 9. In the anther locule of P. armeniaca cultured long in medium, swollen microspores, polynucleate microspores, multicellular pollen grains, or callus mass were frequently observed, this indicating that the callus of this species was microspore-origin. 10. It was clarified that in P. armeniaca production of haploid plant by anther culture might be possible.

• Economic and Environmental Geology
• /
• 제36권3호
• /
• pp.159-170
• /
• 2003

Heavy metals are extracted from Chonju stream sediment, roadside soils and sediments along Honam expressway, soils and tailings from mining area using three different methods (partial extraction in Standard Method, partial extraction method with maintaining 0.1 N of extraction solution and Sequential Extraction Method). In samples having buffer capacity against acid, pH 1 (0.1 N HCl) of extraction solution can not be maintained and pH of extraction solution increases up to 8.0 when partial extraction in Standard Method is used. The averages and ranges of HPE(heavy metals extracted using partial extraction in Standard Method)/HPEM(heavy metals extracted using partial extraction method with maintaining 0.1 N of extraction solution) values are 0.479 and 0.145~0.929 for Cd, 0.534 and 0.078~0.928 for Zn, 0.432 and 0.041~0.992 for Mn, 0.359 and 0.011~0.874 for Cu, 0.150 and 0.018~0.530 for Cr, 0.219 and 0.003~0.853 for Pb, and 0.088 and 1.73${\times}$10$^{-5}$~0.303 for Fe. These data indicate that the difference between HPE and HPEM is large in the order of Fe, Cr, Pb, Cu, Mn, Cd and Zn. The amounts of heavy metals extracted decreases in the follow order; Sum III(sum of fraction I, II, III in sequential extraction)>HPEM>Sum III (sum of fraction I and II)>HPE for Zn, Cd and Mn and Sum III>HPEM>HPE for Cr and Fe. In the case Cr, Sum II is lower than HPEM and higher than HPE. In case of Cu, extracted heavy metals is large in the order Sum IV>HPEM>Sum III HPE. HPE/HPEM value decreases with increasing the amount of HCl used for maintaining 0.1 N of extraction solution. For samples with high buffer capacity, HPE/HPEM value in all elements is lower than 0.2. On the other hand, for samples with low buffer capacity, HPE/HPEM value are over 0.2 and many samples have values higher than 0.6 for Zn, Cd Mn and Cu due to the small difference between Sum II and Sum III, and relatively higher mobility. However, for Fe and Cr, HPE/HPEM value is below 0.2 even for samples with low buffer capacity due to their low mobility and big difference between Sum II and Sum III. This study indicates that the partial extraction method in Korean Standard Method of soil is not suitable for an assessment of soil contamination in area where buffer capacity of soil can be decreased or lost because of a long term exposure to environmental damage such as acidic rain.

• Korean Journal of Poultry Science
• /
• 제29권3호
• /
• pp.195-204
• /
• 2002

Two experiments were conducted to study the effects of nutrient level and feeding method of split diets for a.m. and p.m. on laying hen performance, feed cost and eggshell quality. In experiment 1, 384 ISA Brown layers of 30∼38wk of age were assigned to four treatments which comprise of three replicates each containing 32 birds. The control(C) was fed a conventional single diet throughout the day and split diet groups(T1, T2 and T3) were offered high energy/protein-low Ca diets, and low energy/protein-high Ca diets in a.m.(04:00∼15:00) and p.m.(15:00∼21:00), respectively. In the split diet groups, daily ME and CP consumption, and feed cost were significantly reduced(P<0.05) compared to the C, while the hen-day egg production, average egg weight and daily feed intake were not different among treatments. Due to the reduced daily ME and CP intakes and feed cost, the conversions of feed, ME, CP and feed cost required per day and per kg egg mass were also significantly improved(P<0.05) in the split diet groups. Eggshell qualities (egg specific gravity, egg breaking strength and eggshell thickness) were improved(P<0.05) by split diet feeding. As the Ca level of the p.m. diet increased. In Experiment 2, 384 ISA Brown layers of 50∼58 wk of age were used in three treatments and each treatment was represented by four replicates each containing 32 birds. The control(C) was fed a conventional single diet throughout the day and split diet group(T1) was offered high energy/protein-low Ca diets, and low energy/protein-high Ca diets in a.m.(04:00∼l5:00) and p.m.(15:00∼21:00), respectively. T2 group was fed the diet mixed (50:50) with the a.m. diets in mash and p.m. diet in pellet used T1 group. In T1 and T2 groups, daily feed intake and average egg weight were significantly reduced(P<0.05) compared to the C, while the hen-day egg production was not influenced by the feeding system. Daily ME and CP consumption, and feed cost were reduced(p.0.05), and the conversions of ME, CP and feed cost required per egg were also significantly improved(P<0.05) in T1 and T2, while the conversions of feed, ME, CP and feed cost required per kg egg mass were not different to the C. Eggshell qualities of T1 and T2 were improved(P<0.05) compared to the others. It was concluded the feed and nutrients consumption, feed cost per day or per kg egg mass could be reduced by introducing split diets for a.m. and p.m. and the feeding method of mixed diet of split diets were also convenient and effective for sparing feed cost and improvement of eggshell quality.

• Parasites, Hosts and Diseases
• /
• 제27권4호
• /
• pp.231-248
• /
• 1989

The growth and development of the metacercariae of F. seoulensis cultivated in vitro or on the chick chorioallantois were assessed by comparison with the optimum process of maturation in albino rats and new born chickens. The process of maturation was divided for convenience into six stages: Stage 1 ; cell multiplication, Stage 2; body shaping, Stage 3; separation of genital anlagen, Stage :1 organogeny, Stage 5; gametogony, and Stage 6: oviposition. In Hank's and Tyrode's .solutions, the metacercariae were alive up to 200 days or more at $4^{\circ}C$ without any development. The in vivo maturation process in rats or chicks was as follows: stage 1 from 6 hours; stage 2 from 24 hours; stage 3 from 48 to 72 hours; stage 4 from 3 to 4 days; stage 5 from 4 to 5 days; and stage 6 from 5 to 8 days. Despite unsuccessful infection of the metacercariae to 12 day old chicks, fully mature worms of stage 5 or 6 were recovered from new born chicks (1 to 2 days old), The metacercariae of F. seoulensis grown in vitro were up to stage 3 and no further maturation was observed. Of various media employed, the medium NCTC 109 (Gibco) or NCTC 135(Gibco) supplemented with 20% egg yolk or 20% whole egg macerate or 0.5% yeast was basically required for the earlier development of the fluke. It took 16.1 days(in average) to reach the stage 3 after cultivation. The metacercariae cultivated on the chorioallantoic membranes of 6∼13 day old chick embryo at 37∼38℃ showed their full development up to stage 5 or 6. However, the worms were in general remarkably retarded, compared with those grown in rats or chickens. In the experiments of worm transplant, although the transfer was failed from in vitro culture to in vivo of rats(Per os), the transplants from in vitro culture to the chorioallantois and from the choriollantois to in vivo of rat host were successful with or without development of the transferred worms. In the present study, it was observed that the metacercariae of F, seoulensis can be maintained in vitro media with poor development as well as fully matured in 1 to 2 day-old chicks or on the chorioallantois at a very low rate.