• Journal of the Korea Organic Resources Recycling Association
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• v.18 no.1
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• pp.57-65
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• 2010

Two types of microbial fuel cells(MFC) were continuously operated using synthetic wastewater. One was conventional two-chambered MFC using proton exchange membrane(PEM-MFC), the other was upflow type membraneless MFC(ML-MFC). Graphite felt was used as a anode in PEM-MFC. In membraneless MFC, two MFCs were operated using porous RVC(reticulated vitreous carbon) as a anode. Graphite felt was used as a cathode in all experiments. In experiment of PEM-MFC, the COD removal rate based on the surface area of anode was about $3.0g/m^2{\cdot}d$ regardless of organic loading rate. And the coulombic efficiency amounted to 22.4~23.4%. The acetic acid used as a fuel was transferred through PEM from the anodic chamber to cathodic chamber. The COD removal rate in ML-MFC were $9.3{\sim}10.1g/m^2{\cdot}d$, which indicated the characteristics of anode had no significant effects on COD removal. Coulombic efficiency were 3.6~3.7 % in both cases of ML-MFC experiments, which were relatively small. It was also observed that the microbial growth in cathodic chamber had an adverse effects on the electricity generation in membraneless MFC.

• Applied Biological Chemistry
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• v.21 no.3
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• pp.150-184
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• 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.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 1998.10a
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• pp.83-106
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• 1998

This study was conducted to investigate the probiotics(acid and bile resistance, fermentation properties, viability, cholesterol assimilation, antimicrobial activity, antimutagenicity, and immunoactivation) of the strains of bifidobacteria isolated from healthy Koreans and to investigate the effects of oral administration of Bifidobacterium longum A-2 on the fecal microflora, ${\beta}-glucuronidase$ activity, pH values, Ammonia concentration. The experimental results are summarized as follows: The probiotics were tested for 23 strains including three commer챠al strains as controls. Compared to other strains, strains of A-2 and A-9 showed more acid resistance whereas A-2, A-5, A-13, A-14, A-18 and A-22 showed excellent bile resistances. The properties of bifidobacteria during fermentation were tested. Strains of A-1, A-2, A-3, A-4, A-6, and A-23 resulted in less than pH 4.5 and titratable acidity over 0.90 after 24 hr of fermentation. When the strains of A-2 was grown with glucose, maltose, and fructooligosaccharide, the acetic acid production were higher than with sorbitol and mannitol. The storage stability of the strains of A-2 and A-22 were tesed, indicating the strain A-2 was more stable over 10 days of storage at both $4^{\circ}C$ and $20^{\circ}C$ than A-22. The strains of A-8, A-10, A-11, A-12 and A-20 assimilated more than 30% of cholesterol included in the media. The strains of A-1 and A-2 showed antimicrobial activity against Sta. aureus. The antimutagenicity of the strains were also tested, showing that the mutation was suppressed more by three strains(A-2, A-12, and A-23). In addition, strain A-5 improved immunological activity(phagocytosis, $TNF-{\alpha}$, IL-6) more than other strains. In the effects of oral administration of Bif. longum A-2, the number of fecal bifidobacteria was siginificantly increased(p<0.01) and the level of fecal ${\beta}-glucuronidase$ also was siginificantly reduced(p<0.05). However there were no siginificant differences in the level of Lαctobacilli, Enterobacteriaceae, Clostridium perfringens, pH and ammonia by the administration. The results suggested that Bif. longum A-2 may be met the criteria for probiotics culture.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.6
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• pp.836-842
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• 2009

The objectives of this study were to evaluate potential benefits of intercropping of corn with runner bean for a smallsized farming system, based on land equivalent ratio (LER) and silage yield and quality of corn intercropped with runner bean (Phaseolus vulgaris L.), in arid conditions of Turkey under an irrigation system. This experiment was established as a split-plot design in a randomized complete block, with three replications and carried out over two (consecutive) years in 2006 and 2007. Seven different mixtures (runner bean, B and silage corn sole crop, C, 10% B+90% C, 20% B+80% C, 30% B+70% C, 40% B+60%C, and 50% B+50%C) of silage corn-runner bean were intercropped. All of the mixtures were grown under irrigation. The corn-runner bean fields were planted in the second week of May and harvested in the first week of September in both years. Green beans were harvested three times each year and green bean yields were recorded each time. After the 3rd harvest of green bean, residues of bean and corn together were randomly harvested from a 1 $m^{2}$ area by hand using a clipper when the bean started to dry and corn was at the dough stage. Green mass yields of each plot were recorded. Silages were prepared from each plot (triplicate) in 1 L mini-silos. After 60 d ensiling, subsamples were taken from this material for determination of dry matter (DM), pH, organic acids, chemical composition, and in vitro DM digestibility of silages. The LER index was also calculated to evaluate intercrop efficiencies with respect to sole crops. Average pH, acetic, propionic and butyric acid concentrations were similar but lactic acid and ammonia-N levels were significantly different (p<0.05) among different mixtures of bean intercropped with corn. Ammonia-N levels linearly increased from 0.90% to 2.218 as the percentage of bean increased in the mixtures up to a 50:50 seeding ratio. While average CP content increased linearly from 6.47 to 12.45%, and average NDF and ADF contents decreased linearly from 56.17 to 44.88 and from 34.92 to 33.51%, respectively, (p<0.05) as the percentage of bean increased in the mixtures up to a 50:50 seeding ratio, but DM and OM contents did not differ among different mixtures of bean intercropped with corn (p>0.05). In vitro OM digestibility values differed significantly among bean-corn mixture silages (p<0.05). Fresh bean, herbage DM, IVOMD, ME yields, and LER index were significantly influenced by percentage of bean in the mixtures (p<0.01). As the percentage of bean increased in the mixtures up to a 50:50 seeding ratio, yields of fresh bean (from 0 to 24,380 kg/ha) and CP (from 1,258.0 to 1,563.0 kg/ha) and LER values (from 1.0 to 1.775) linearly increased, but yields of herbage DM (from 19,670 to 12,550 kg/ha), IVOMD (from 12,790 to 8,020 kg/ha) and ME (46,230 to 29,000 Mcal/ha) yields decreased (p<0.05). In conclusion, all of the bean-corn mixtures provided a good silage and better CP concentrations. Even though forage yields decreased, the LER index linearly increased as the percentage of bean increased in the mixture up to a 50:50 seeding ratio, which indicates a greater utilization of land. Therefore, a 50:50 seeding ratio seemed to be best for optimal utilization of land in this study and to provide greater financial stability for labor-intensive, small farmers.

• Korean Journal of Food Science and Technology
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• v.30 no.2
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• pp.425-433
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• 1998

The characteristics of the reaction of calcium carbonate $(CaCO_3)$ immobilized with alginate as buffer system for the high concentration cultivation of bifidobacteria in fermenter are described by the mathematical model, and tested for the reusing possibility of the used $CaCO_3$ beads. When$CaCO_3$ beads with the various diameters were reacted in 0.1 M of the mixed organic acids (0.6 M of acetic acid and 0.4 M lactic acid) and in fermenter inoculated Bifidobacterium longum ATCC 15707, the change of bead diameters can be calculated with the amount of the decreased $CaCO_3$ from the surface of bead using the mathematical model. These values was similar to the directly measured bead diameter by a micrometer. Therefore, it was considered that the mathematical model could be used for explaining the reaction charateristics of the $CaCO_3$ bead reacted with the organic acids. When Bifidobacterium longum was incubated at $37^{\circ}C$ for 20 hours in fermenter with $CaCO_3$ beads, the buffering effect of $CaCO_3$, the reduce rate of the bead diameter, and the growth rate of Bifidobacterium longum were higher at the smaller beads than beads with the larger diameters. Also, when Bifidobacterium longum was incubated in fermenter with the mixed beads which were added new beads to the recovered beads in order to equalize with the total surface area of initial beads, the buffering effect of $CaCO_3$ bead and the growth rate of Bifidobacterium longum were very corresponded with the results of the fermentation using the only initial beads. Therfore, it is expected that the used beads can be reused by adding the initial beads.

• Journal of Food Hygiene and Safety
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• v.26 no.1
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• pp.64-69
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• 2011

Steamed rice is usually used as an essential ingredient when Takju or Yakju is brewed in Korea. Alternatively, non-steamed rice can be used to keep thermolabile nutrients and fresh tastes richer in Takju or Yakju. In this study, therefore, the physicochemical properties (ethanol and sugar contents, pH, total acidities, and turbidities) and the fermentative microbial profiles (aerobic mesophillic bacteria (AMB), fungi, lactic acid bacteria (LAB), acetic acid bacteria (AAB), and Escherichia coli and coliforms) have been compared among 4 Takju and 1 Yakju samples brewed using steamed or non-steamed rice. Yakju brewed using non-steamed rice has approximately 2-3 times higher ethanol and sugar contents than other 4 Takjus brewed using steamed or non-steamed rice. The pH and total acidity values of all the 5 samples ranged 3.77-4.30 and 0.12-0.35, respectively. As for turbidities, Yakju brewed using nons-teamed rice was transparent, but other 4 Takjus were not. The AMB and fungal counts for Yakju brewed using nons-teamed rice were approximately $10^4$-fold less than those for 4 Takjus. The LAB counts for Takju and Yakju brewed using non-steamed rice were $10^3$-fold less than those for Takjus brewed using steamed rice. The AAB counts ranged $2-6\;log_{10}\;CFU/mL$ for all the 5 samples. E.coli and coliforms were not detected. Overall, there was no significant difference in microbial counts among 4 Takjus brewed using steamed or non-steamed rice, but Takju has higher microbial counts than Yakju. All the 5 samples were conclusively considered to be hygienically brewed and processed containing plenty of beneficial microorganisms.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.5
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• pp.563-569
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• 2001

In order to utilize the processing wastes of squid, chitosan was prepared by intermittent deacetylation treaoent of $\beta-chitin$ contained richly in the pen of squid. Acetylchitosan also was synthesized from squid pen chitosan with anhydrous acetic acid and their characteristics were investigated. The amounts of nitrogen and ash of squid pen chitosan were $5.80.2\% and 0.2\pm0.03\%$ respectively, the yield of squid pen chitosan was $25\pm3\%$, the degree of deacetylation was $92\%$, and the molecular weight was $1.15\times10^6$, Acetyl contents of N-acetylchitosan powder, acetylchitosan bead, N-ACF-1 (N-acetylchitosan film-1) and N-ACF-2 (N-acetylchitosan film-2) were $55.9\%, 63.2\%, 56\% and 58.7\%$ respectively. Two major peaks, amide I ($1,653 cm^{-1}$) and II ($1,558 cm^{-1}$) bent, on FT-IR spectra of the N-acetylchitosan from squid pen were almost similar to these of $\beta-chitin$, While there was a broad single peak at $1,601 cm^{-1}$assigned to be an amide I bend in squid pen chitosan. The CP/MAS NMR spectra of $\beta-chitin$, squid pen chitosan and N-acetylchitosan from squid pen showed a relative broad and single peak at 74 ppm assigned to fifth carbon (C-5) and third carbon (C-3). In case of $\beta-chitin$ and N-acetylchitosan from squid pen, single peak at 74 ppm was showed as the same of $\beta-chitin$ type.

• Korean Journal of Microbiology
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• v.17 no.3
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• pp.97-130
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• 1979

Many industrial processes those employ bacteria are subjected to phage infestations. In L-glutamic acid fermentions using acetic acid, the phage infestations of the organisms have been recently recognized. In efforts to elucidate the sources of phage contamination involved in the abnormal fermentation, a series of study was conducted to isolate the phages both from the contents of abnormally fermented tanks and the soil or sewage samples from the surroundings of a fermentation factory, to define major charateristics of the phage isolates, and finally to determine the correlation between the phage isolates and temperate phages originating from the miscellaneous bacterial species isolated from the soil or sewage samples. The results are summarized as follows; 1) All phages were isolated from the irregular fermentation tanks and soil or sewage samples, and they were designated as phage PR-1, PR-2, PR-3, PR-4, PR-5, PR-6, and PR-7, in the order of isolation. These PR-series phages were proved to be highly specific for the variant strains of Br. lactofermentum only, namely, phage PR-1 and PR-2 for Br. lactofermentum No. 468-5 and phage PR-3~PR-7 for Br. lactofemrentum No. 2256. By cross-neutralization test, the 7 phagescould be subdivided into 3 groups, i. e., phage PR-I and PR-2 the first, phage PR-3, PR-4, PR-5, PR-6 the second, and the phage PR-7 the third. 2) The 7 phages were virulent under the experimental conditions. They produced plaques with clear and relatively sharp margins without distinct halo. The mean sizes of plaques were 1.5mm in diameter for phage PR-1 and PR-2, and 1. Omm for phages PR-3~PR-7. Double layer technique modified by Hongo and described by Adams, was applied to assay of the PR-series phages. The factors influencing the plaques were as follows;young age cells of host bacteria cultured for 3-6 hours represented the largest number and size, optimum was pH 7.0, incubation temperature was $30^{\circ}C$, and agar concentration and amount of overlayer medium were 0.6% and 0.2ml, respectively. 3) PR-series phages were stable in 0.05M tris buffer and 0.1M ammonium acetate buffer solution. The addition of $5{\times}10^{-3}M$ magnesium ion effectively increased the stability. Thermostability experiments indicated that PR-series phages were stable at the teinperture between $50^{\circ}{\sim}55^{\circ}C$ in nutrient medium, $45^{\circ}{\sim}50^{\circ}C$ in buffer solution. However, the phages mere completely inactivated at 603C and 65$^{\circ}$C within 10 minutes. The phages were stable at the range of pH6~9 in nutrient medium and of pH 8-9 in buffer solution, respectively. Exposure of the phages to UV for 25, 60 and 100 seconds resulted in the complete loss of infectivily, respectively. 4) Electron microscopy showed that PR-series phage particles exhibited rather similar morphology, differing in the size All of PR-series phages had a multilateral head and had a simple long tiil about three to five times long as compared with head. By the size, phage PR-1 and PR-2, PR-3, PR-4, PR-5, and PR-6 and PR-7 were classified into same groups, respectively. The head and tail size of phage PR-1, PR-5, PR-5(T) and PR-7 were 85nm, 74nm and 235nm and 350mm, and 72nm and 210nm, respectively. 5) Nucleic acids of PR-series phages were double stranded DNA. The G+C contents of phage PR-1, PR-5 and PR-7 were 56.1, 52.9 and 53.7, respectively. The values of G+C contents derived from the $T_m$ were in agreement with the chemically determined values. 6) PR-series phages effectively adsorbed on their host bacteria at the rate of more than 90% during 5 min. K value for phage PR-1, PR-5 and PR-7 were calculated to be $6{\times}10^9 ml$ per minute, respectiveky. The pH of the medium did effect adsorption rate, but both temperature and age of host cells did not. Generally, optimum adsorption condition of phages seemed to be almost same as optimum growth conditions of host bacteria. 7) In one-step growth experiments, the latent periods at $30^{\circ}C$ for PR-1, and PR-7 were about 70, 50 and 55 min, respectively. The corresponding average burst size was 200, 70 and 90, respectively. Lpsis period according to the multiplicity of infection and a phage series. In case of m. o. i. 100, strain No. 2256 (PR-5) and No. 468-5(PR-1) failed to grow and turbidity decreased after 50 and 70min, respectively. 8) In the lysate of a plaque purified phage PR-5 infected bacteria, there observed 2 types ofphage particles, i. e., phage PR-5 and PR-5 (T) of similar morphology but differing at the length of phage tail, and phage tail like particles. The phage taillike particles could be divided into 4 types by the length. Induction experiments of Br. lactofermentum with UV irradiation, mitomycin C or bacitracin treatment produced neither phage PR-5 (T) or phage tail-like particles. 9) No lysis occured when the growth of 7 strains of miscellaneous bacteria, isolated from soil and sewage samples, were inoculated with either phage PR-5 (T) or phage tail-like particles the inoculation of phage PR-5 pellet resulted in the growth inhibition of the orgainsms in the spot test. The lysates obtained from 3 miscellaneous soil derived bacteria following mitomycin C treatment the growth of Br. lactofermentum, but did not lyze the bacterium.

• Journal of Korean Society of Forest Science
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• v.82 no.1
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• pp.26-33
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• 1993

In vitro shoot proliferation and rooting were tested for 2-0 seedlings of half-sib families of 4 plus oaks trees. Nodal segments having axillary buds from 37 families(16 of Quercus acutissima, 10 of Q. variabilis, 7 of Q. serrata, and 4 of Q. mongolica) were cultured on WPM(Woody Plant Medium) supplemented with 0.5 mg/l BA (6-benzyladenine) and 0.01 mg/l NAA(${\alpha}$-naphthalene acetic acid) and subcultured at 2-3 weeks of intervals fur 6 months. In vitro rooting was carried out on GD(Gresshoff and Doy) medium supplemented with 0.5mg/l IBA(indole butyric acid). The capacity for shoot proliferation and rooting was highly varied with families. Generally, white oaks(Q. serrata and Q. mongolica) showed poor response than black oaks(Q. acutissima and Q, variabilis) in shoot proliferation and rooting. Among the total of 37 families, 7 of Q. acutissima, each 2 of Q. variabilis, Q. serrata, and Q. mongolica revealed abilities for continuous shoot proliferation, and the others failed to proliferate. Rooting of the selected oak trees also greatly varied among the families. In Q. acutissima, rooting ratio ranged from 10.0%(CB 25. KG 4) to 89.8%(CB 18). Although 26.7% of KG 16 in Q. variabilis, 3.3% of JN 15 in Q. serrata were rooted, Q. mongolica was not rooted at all in this experimental conditions. No relationship between shoot growth and the rooting ability was observed. Present results suggest the possibility of large-scale micropropagation, but further studies on family differences, shoot-tip necrosis, and callusing of rooting junction are still required to develop reliable micropropagation systems.

• Korean Journal of Organic Agriculture
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• v.25 no.4
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• pp.901-916
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• 2017

This study was conducted to evaluate natural plant extracts for methane gas reduction in ruminants. Rumen fluid was collected from cannulated Hanwoo cow ($450{\pm}30kg$) consuming 400 g/kg concentrate and 600 g/kg timothy. The 15 ml of mixture comparing McDougall's buffer and rumen fluid in the ratio 2 to 1, was dispensed anaerobically into 50 ml serum bottles. Rumen fluid contents were collected and in vitro fermentation prepared control (timothy, 300 mg), ginseng, balloon flower, yucca plant, camellia, tea plant and ogapi extracts were added at the level of 5% against 300 mg of timothy as a substrate (v/w) and incubated for 3, 6, 9, 12, 24, 48, and 72 h. In vitro pH values range 6.55~7.41, this range include rumen titration. The dry matter digestibility was not differ between all treatments and control. Total gas emission was significantly higher (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Carbon dioxide emission was not differ all treatments on 9 h than in control and significantly higher (p<0.05) yucca plant, camellia and tea plant treatments on 12 h than control. Methane emission was not differ all treatments on 6 h than in control. The rumen microbial growth rate was significantly higher (p<0.05) in ginseng, balloon flower on 12 h and significantly higher (p<0.05) in ginseng, yucca plant, tea plant and ogapi treatments on 24 h than in control. Total VFA was significantly higher (p<0.05) in tea plant and ogapi treatments on 12 h than in control and significantly higher (p<0.05) in ginseng, balloon flower treatments on 48 h than in control. Acetic acid was significantly lower (p<0.05) in ginseng and balloon flower treatments on 24 h than in control. Propionic acid was significantly higher (p<0.05) in ginseng and balloon flower treatments on 48 h than in control. As a results, sixth natural plant extracts had no significant effect dry matter digestibility and negative on rumen fermentation, but not effect methane reduction.