• Economic and Environmental Geology
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• v.31 no.3
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• pp.201-213
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• 1998

From the Jungwon and Munkyeong areas which are among the famous producers of the carbonate-type groundwaters in Korea, various kinds of natural waters (deep groundwater, shallow groundwater and surface water) were collected between 1996 and 1997 and were studied for hydrogeochemical and environmental isotope (${\delta}^{34}S_{so4}$, ${\delta}^{18}O$, ${\delta}D$)systematics. Two types of deep groundwaters (carbonate type and alkali type) occur together in the two areas, and each shows distinct hydrogeochemical and environmental isotope characteristics. The carbonate type waters show the hydrochemical feature of the 'calcium(-sodium)-bicarbonate(-sulfate) type', whereas the alkali type water of the 'sodium-bicarbonate type'. The former type waters are characterized by lower pH, higher Eh, and higher amounts of dissolved ions (especialJy, $Ca^{2+}$, $Na^{+}$, $Mg^{2+}$, $HCO_3{^-}$ and $SO_4{^{2-}}$). Two types of deep groundwaters are all saturated or supersaturated with respect to calcite. Two types of deep groundwaters were both derived from pre-thermonuclear (about more than 40 years old) meteoric waters (with lighter 0 and H isotope data than younger waters, i.e., shallow cold groundwaters and surface waters) which evolved through prolonged water-rock interaction. Based on the geologic setting, water chemistry, and environmental isotope data, however, each of these two different types of deep groundwaters represents distinct hydrologic and hydrogeochemical evolution at depths. The carbonate type groundwaters were formed through mixing with acidic waters that were derived from dissolution of pyrites in hydrothermal vein ores (for the Jungwon area water) or in anthracite coal beds (for the Munkyeong area water). If the deeply percolating meteoric waters did not meet pyrites during the circulation, only the alkali type groundwaters would form. This hydrologic and hydrogeochemical model may be successfully applied to the other carbonate type groundwaters in Korea.

• Applied Biological Chemistry
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• v.16 no.1
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• pp.1-11
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• 1973

In order to obtain the basic informations on the production of single cell protein from ethanol, 145 yeast strains utilizing ethanol as a sole carbon source were isolated from 32 soil samples in Korea. A yeast strain showing the highest cell yield among the isolated strains was selected and identified. The optimum culture condition, utilization of other carbon sources and the cultural characteristics for the selected yeast, and the chemical analysis of the yeast cell composition, and utilization of ethanol by the selected yeast were investigated. All the culture was carried out in the shaking flasks. The results obtained were as follows: 1. The selected yeast strain was identified as Debaryomyces nicotianae-SNU 72. 2. The optimum composition of the medium for the selected yeast is : Ethanol 40 ml, Urea 0.5 g, Potassium phosphate (dibasic) 0.5 g, Ammoium phosphate (monobasic) 0.15 g, Magnesium sulfate 0.05 g, Calcium chloride 0.01g, Yeast extract 0.005 g, Tap water 1000 ml. 3. The optimum pH was 5.0-5.5, the optimum temperature $30-33^{\circ}C$ and the aerobic state was unimportant. 4. Utilization of methanol, n-propanol, iso-propanol, n-butanol, iso-butanol, tert-amyl alcohol and acetic acid by the selected yeast was very weak. So substitution of the subtrate was thought to be impossible. 5. Studies on the propagation of the yeast cells showed that the lag phase of the yeast cells lasted 16 hours, and the logarithmic growth phase extended 16 to 28 hours. The specific growth rate was about $0.19\;hr^{-1}$ and the doubling time was 3.6 hours during the logarithmic growth phase. 6. As the result of the chemical analysis of the dry yeast cells, the content rate of the crude protein was 55.19 %, the content of others was similar to the average content of the yeast component. 7. After 34 hours cultivation, under the optimum culture condition investigated, the dry cell yield against the amount of the added ethanol was 53.4 % (W/V%), the dry cell yield against the amount of the utilized ethanol was 73.6 % (W/V%), the evaporation rate of ethanol was about 19.1 %.

• Conservation Science in Museum
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• v.30
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• pp.23-46
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• 2023

The Jija Chongtong Gun, owned by Seokdang Museum of Dong-A University, is a tubedstyle heavy weapon of the battlefield in the mid-Joseon Dynasty and is the second largest firearm after Cheonja Chongtong. The original surface color of the Jija Chongtong Gun was obscured by foreign substances and therefore it was judged that its condition requires the conservation treatment. For stable conservation treatment, gamma ray and X-ray non-destructive transmission surveys was conducted to determine the internal structure and conservation condition. And the component analysis on the material components and surface contaminants of Jija Chongtong Gun was conducted by utilizing the p-XRF component analysis, SEM-EDS component analysis, and XRD analysis. As a result of the gamma-ray and X-ray non-destructive transmission investigation, a large amount of air bubbles was observed inside Jija Chongtong Gun, and the part that appeared to be a chaplet by visual observation was not identified. As a result of gamma-ray and p-XRF component analysis, it was confirmed that Jija Chongtong Gun was bronze made of copper (Cu), tin (Sn), and lead (Pb) alloy. As a result of surface analysis of foreign substances using SEM-EDS, it was confirmed that the main components of white foreign substances were calcium (Ca), sulfur (S), and titanium (Ti). Titanium was presumed to be titanium dioxide (TiO₂), the main component of white correction fluid. The red foreign substance was confirmed to contain barium (Ba) as its main ingredient, and was presumed to be barium sulfate (BaSO₄), an extender pigment in paint. White and red contaminants, mainly composed of titanium and barium, are presumed to have been deposited on the surface in recent years. The yellow foreign substances were confirmed to be aluminum (Al) and silicon (Si), and were presumed to have originated from soil components. As a result of SEM-EDS and XRD component analysis, the white foreign substance was confirmed to be gypsum (CaS). Based on the results of component analysis, surface impurities were removed, stabilization treatment, and strengthening treatment were performed. During the conservation process, unknown inscriptions Woo (右), Byeong (兵), Sang (上), and Yi (二) were discovered through a portable microscope and precise 3D scanning. In addition, the carving method, depth, and width of the inscription were measured. Woo Byeong Sang is located above Happo Fortress in Changwon, and Yi can be identified as the second hill.

• 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.

• Applied Biological Chemistry
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• v.29 no.1
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• pp.62-72
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• 1986

At present, the definition and chemical analysis method of available soil phosphorus for plants have not been standardized because of the complexity of crop and soil characteristics in Korea and many analysis methods have been suggested with different extraction conditions. Suitable analytical method of available soil P should be established by the trial of various methods based on crop nutrition and soil conditions. To establish the most suitable analysis method of available soiIP, a pot experiment with young maize was conducted over 44 different upland soils collected over the land of Korea. The amount of uptaken P by the plant was determined by ten different chemical methods for the available soil P. The results obtained were as follows: 1. Total phosphorus content in the sample soils ranged ranged $533{\sim}4917\;ppm$, and showed significant positive correlation with the content of organic matter. 2. The P content was relatively low in the acid sulfate soil and very high in the volcanic ash soil although both types of soil contained high level of orgic matter. 3. The amount of extractable P determined by ten different methods were varied more or less, and the ratios of the extractable P to the total soil P were in the range of $1{\sim}48%$. 4. The relative values to the amount of extractable soil P by different methods were in the order of $H_2O(5\;min.)\;1.0\;<\;H_2O(60min.)\;2.27\;<\;NH_4HCO_3\;5.57\;<\;NaHCO_3\;7.42\;<\;Double\;lactate\;9.71\;<\;Bray\;No.1\;12.53\;<\;Lancaster\;17.63\;<\;Nelson\;25.96\;<\;AcOH\;27.6\;<\;CAL-method\;50.27$ 5. The amount of extractable P determined by all of applied methods was very low in acid sulfate soil, volcanic ash soil and coarse textured soil. 6. Soil pH and total soil P generally showed significant positive correlation with the chemically extracted P, and soil organic matter was negatively correlated with the determined by Nelson-and CAL-method. Olsen method which showed significant correlation with exchangeable calcium seemed to be recommendable for calcareous soils. 7. Total amount of uptaken P by Young maize through continuos twice cropping was 4.05% of total soil P in average, and the uptake in the second cropping was twice as much as that of the first cropping. 8. Three determination methods, i.e. Soltanpour-, Double lactate and Bray No. 1-method seemed to be more suitable than Lancaster method which is widely practiced at present in Korea. However, further study should be carried out with other crops and soils to most adequate chemical method for determination of available soil P.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.14 no.2
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• pp.47-58
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• 1981

Relationships between the electrical conductivity and the contents of the chloride, sulfate, calcium, magnesium, sodium, potassium and total major inorganic ions, and between each, chemical conservative constituents were calculated with the data which sampled at the lesions of Mulgeum and between Namji and Wondong from March 1974 to April 1980. Semilogarithmic relations were found between the electrical conductivity and the contents of monovalent ions, and logarithmic relations were found between the electrical conductivity and the contents of divalent ions at the both regions. The relational equations between the electrical conductivity $\lambda_{25}$and the contents of the major inorganic ions at Mulgeum are as follows: $log\;Cl(ppm)\;=\;2.37{\cdot}\lambda_{25}(m{\mho}/cm)+0.733{\pm}0.141$, $log\;SO_4(ppm)=1.12{\cdot}log\lambda_{25}(m{\mho}/cm)+2.14{\pm}0.18$, $log\;Ca(ppm)=0.615{\cdot}log\lambda_{25}(m{\mho}/cm)+1.67{\pm}0.12$, $log\;Mg(ppm)=0.756{\cdot}log\lambda_{25}(m{\mho}/cm)+1.27{\pm}0.11$, $log\;Na(ppm)=2.82{\cdot}\lambda_{25}(m{\mho}/cm)+0.551{\pm}0.133$, $log\;K(ppm)=1.33{\cdot}\lambda_{25}(m{\mho}/cm)+0.136{\pm}0.095$, and total inorganic ions $C(ppm)=399{\cdot}\lambda_{25}(m{\mho}/cm)-0.9{\pm}14.6$. The relational equations between the electrical conductivity ($\lambda_{25}$) and the contents of the major inorganic ions at the region between Namji and Wondong a.e as follows: $log\;Cl(ppm)=4.27{\cdot}\lambda_{25}(m{\mho}/cm)+0.380{\pm}0.138$, $log\;SO_4(ppm)=0.915{\cdot}log\lambda_{25}(m{\mho}/cm)+1.95{\pm}0.18$, $log\;Ca(ppm)=0.756{\cdot}log\lambda_{25}(m{\mho}/cm)+1.74{\pm}0.12$, $log\;Mg(ppm)=1.00{\cdot}log\lambda_{25}(m{\mho}/cm)+1.41{\pm}0.10$. $log\;Na(ppm)=2.47{\cdot}\lambda_{25}(m{\mho}/cm)+0.614{\pm}0.065$, $log\;K(ppm)=1.62{\cdot}\lambda_{25}(m{\mho}/cm)+0.030{\pm}0.060$, and total inorganic ions $C(ppm)=323{\cdot}\lambda_{25}(m{\mho}/cm)+11.7{\pm}9.3$. Logarithmic relations were found between each chemical conservative constituents at Mulgeum and the equations are as follows: $log\;Cl(ppm)=0.711{\cdot}log\;SO_4(ppm)+0.488{\pm}0.206$, $log\;Cl(ppm)=0.337{\cdot}log\;Ca(ppm)+0.822{\pm}0.130$, $log\;Cl(ppm)=0.605{\cdot}log\;Mg(ppm)-0.017{\pm}0.154$, $Cl(ppm)=0.676{\cdot}Na(ppm)+2.31{\pm}4.67$, $log\;Cl(ppm)=0.406{\cdot}log\;K(ppm)-0.092{\pm}0.112$, $log\;SO_4(ppm)=0.378{\cdot}log\;Ca(ppm)+0.721{\pm}0.125$, $log\;SO_4(ppm)=0.462{\cdot}log\;Mg(ppm)+0.107{\pm}0.118$, $log\;SO_4(ppm)=0.592{\cdot}log\;Na(ppm)+0.313{\pm}0.191$, $log\;SO_4(ppm)=0.308{\cdot}log\;K(ppm)-0.019{\pm}0.120$, $Ca(ppm)=0.262{\cdot}Mg(ppm)+0.74{\pm}1.71$. $log\;Ca(ppm)=1.10{\cdot}log\;Na(ppm)-0.243{\pm}0.239$, $Ca(ppm)=0.0737{\cdot}K(ppm)+1.26{\pm}0.73$, $log\;Mg(ppm)=0.0950{\cdot}Na(ppm)+0.587{\pm}0.159$, $log\;Mg(ppm)=0.0518{\cdot}K(ppm)+0.111{\pm}0.102$, and $Na(ppm)=0.0771{\cdot}K(ppm)+1.49{\pm}0.59$. Logarithmic relations were found between each chemical conservative constituents except a relationship between the chloride and calcium contents at the region between Namji and Wondong, and the equations are as follows : $log\;Cl(ppm)=0.312{\cdot}log\;SO_4(ppm)+0.907{\pm}0.210$, $log\;Cl(ppm)=0.458{\cdot}log\;Mg(ppm)+0.135{\pm}0.130$, $Cl(ppm)=0.484{\cdot}logNa(ppm)+0.507{\pm}0.081$, $Cl(ppm)=0.0476{\cdot}K(ppm)+1.41{\pm}0.34$, $log\;SO_4(ppm)=0.886{\cdot}log\;Ca(ppm)+0.046{\pm}0.050$, $log\;SO_4(ppm)=0.422{\cdot}log\;Mg(ppm)+0.139{\pm}0.161$, $log\;SO_4(ppm)=0.374{\cdot}log\;Na(ppm)+0.603{\pm}0.140$, $log\;SO_4(ppm)=0.245{\cdot}log\;K(ppm)+0.023{\pm}0.102$, $log\;Ca(ppm)=0.587{\cdot}log\;Mg(ppm)+0.003{\pm}0.088$, $log\;Ca(ppm)=0.892{\cdot}log\;Na(ppm)+0.028{\pm}0.109$, $log\;Ca(ppm)=0.294{\cdot}log\;K(ppm)-0.001{\pm}0.085$, $log\;Mg(ppm)=0.600{\cdot}log\;Na(ppm)+0.674{\pm}0.120$, $log\;Mg(ppm)=0.440{\cdot}log\;K(ppm)+0.038{\pm}0.081$, and $log\;Na(ppm)=0.522{\cdot}log\;K(ppm)-0.260{\pm}0.072$.

• The Korean Journal of Mycology
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• v.7 no.1
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• pp.13-73
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• 1979

These studies were conducted to investigate nutrient sources and supplementary materials of synthetic compost media for Agaricus bisporus culture. Investigation were carried out to establish the optimum composition for compost of Agaricus bisporus methods of out-door fermentation and peakheating with rice straw as the main substrate of the media. The incidence and flora of harmful organisms in rice straw compost and their control were also studied. 1. When rice straw was used as the main substrate in synthetic compost as a carbon source. yields were remarkably high. Fermentation was more rapid than that of barley straw or wheat straw, and the total nitrogen content was high in rice straw compost. 2. Since the morphological and physico-chemical nature of Japonica and Indica types of rice straw are greatly dissimilar. there were apparent differences in the process of compost fermentation. Fermentation of Indica type straw proceeded more rapidly with a shortening the compost period, reducing the water supply, and required adding of supplementary materials for producing stable physical conditions. 3. Use of barley straw compost resulted in a smaller crop compared with rice straw. but when a 50%, barley straw and 50% rice straw mixture was used, the yield was almost the same as that using only rice straw. 4. There were extremely high positive correlations between yield of Agaricus bisporus and the total nitrogen, organic nitrogen, amino acids, amides and amino sugar nitrogen content of compost. The mycerial growth and fruit body formation were severely inhibited by ammonium nitrogen. 5. When rice straw was used as the main substrate for compost media, urea was the most suitable source of nitrogen. Poor results were obtained with calcium cyanamide and ammonium sulfate. When urea was applied three separate times, nitrogen loss during composting was decreased and the total nitrogen content of compost was increased. 6. The supplementation of organic nutrient activated compost fermentation and increased yield of Agaricus bisporus. The best sources of organic nutrients were: perilla meal, sesame meal, wheat bran and poultry manure, etc. 7. Soybean meal, tobacco powder and glutamic acid fermentation by-products which were industrial wastes, could be substituted for perilla meal, sesame meal and wheat bran as organic nutrient sources for compost media. B. When gypsum and zeolite were added to rice straw. physical deterioration of compost due to excess moisture and caramelization was observed. The Indica type of straw was more remarkable in increase of yield of Agricus bisporus by addition of supplementing materials than Japonica straw. 9. For preparing rice straw compost, the best mixture was prepared by 10% poultry manure, 5% perilla meal, 1. 2 to 1. 5% urea and 1% gypsum. At spring cropping, it was good to add rice bran to accelerate heat generation of the compost heap. 10. There was significantly high positive correlation (r=0.97) between accumulated temperature and the decomposition degree of compost during outdoor composting. The yield was highest at accumulated temperatures between 900 and $1,000^{\circ}C$. 11. Prolonging the composting period brought about an increase in decomposition degree and total nitrogen content, but a decrease in ammonium nitrogen. In the spring the suitable period of composting was 20 to 25 days. and about 15 days in autumn. For those periods, the degree of decomposition was 19 to 24%. 12. Compactness of wet compost at filling caused an increase in the residual ammonium nitrogen. methane and organic acid during peak heating. There was negative correlation between methane content and yield (r=0.76)and the same was true between volatile organic acid and yield (r=0.73). 13. In compost with a moisture content range between 69 to 80% at filling. the higher the moisture content, the lower the yield (r=0.78). This result was attributed to a reduction in the porosity of compost at filling the beds. The optimum porosity for good fermentation was between 41 and 53%. 14. Peak heating of the compost was essential for the prevention of harmful microorganisms and insect pests. and for the removal of excess ammonia. It was necessary to continue fer mentatiion for four days after peak heating. 15. Ten species of fungi which are harmful or competitive to Agaricus bisporus were identified from the rice compost, including Diehliomyces microsporus, Trichoderma sp. and Stysanus stemoites. The frequency of occurrance was notably high with serious damage to Agaricus bisporus. 16. Diehliomyces microsporus could be controlled by temperature adjustment of the growing room and by fumigating the compost and the house with Basamid and Vapam. Trichoderma was prevented by the use of Bavistin and Benomyl. 17. Four species of nematodes and five species of mites occured in compost during out-door composting. These orgnanisms could be controlled through peakheating compost for 6 hours at $60^{\circ}C$.