• Tuberculosis and Respiratory Diseases
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• v.59 no.4
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• pp.389-396
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• 2005

Background : Central venous catheters(CVCs) area major source of nosocomial infection. Chlorhexidine-silver sulfadiazine coated catheters (CHSS) were developed to reduce the rate of CVC infection. However, the clinical effectiveness of CHSS in comparison wth non-coated catheter (NCC) remains to be evaluated. Methods : From January 2004 to December 2004 in medical intensive care unit (ICU) of Asan Medical Center, CVCs were inserted in 446 cases. We retrospectively analyzed characteristics of patients and catheterization,the catheter-related infection rate and colonization, microbiologic findings, and insertion sites (subclavian, jugular, femoral) according to the type of inserted CVCs (NCC: 187 cases, CHSS: 259 cases). Catheter related infection is defined as catheter related bacteremia and catheter related non-bacteremic sepsis. Results : 1) The mean age of the patients in each group was $62{\pm}16$ years, $63{\pm}15$ years (p=0.42), and sex ratio 94:50, 141:69 (p=0.9) in NCC and CHSS. Duration of ICU admission ($29{\pm}37$, $26{\pm}44$ p=0.42), duration of mechanical ventilation ($17{\pm}22$, $15{\pm}19$ p=0.17), and APACHE III score at the time of CVC insertion ($81{\pm}34$, $82{\pm}37$ p=0.61) were not different between both groups. 2) Mean duration of catheterization was 118 in NCC and 119 in CHSS (p=0.98). Number of catheter-days was 2176 days in NCC and 3035 days in CHSS. Catheter-related infection occurred in 9 (4.8%) cases receiving NCC and 4 cases (1.5%) receiving CHSS. Catheterrelated infection incidence per 1000 catheter-days was 4.1 and 1.3, respectively (p=0.04). CHSS was associated with a significant reduction of infection in jugular catheters regarding to insertion sites (p=0.01). 3) Microorganisms causing infection were Staphylococcus aureus (n=3), Candida (n=3), coagulase-negative Staphylococci (n=2), and Klebsiella (n=1) in NCC, and Candida species (n=2), coagulase-negative Staphylococci (n=2), Proteus (n=1) in CHSS. Conclusion : CHSS has significantly reduced the episodes of infection compared to NCC in jugular catheterization in medical ICU.

• Journal of Life Science
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• v.19 no.11
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• pp.1538-1546
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• 2009

This study was conducted to estimate the in vitro fermentation characteristics and in situ degradabilities of total mixed rations fermented by the synbiotic co-cultures composed of various anaerobic microorganisms in the rumen of cow. Seventy two TMR bags (4 treatments $\times$ 6 fermentation days $\times$ 3 replications) were manufactured for in vitro and in situ experiments. The experiment was composed of four treatments including the control, the mould and bacteria synbiotics (T1), the mould and yeast synbiotics (T2) and the bacteria and yeast synbiotics (T3). Each treatment had six fermentation days (1, 3, 5, 7, 14, 21 day) with three replications. Two rumen cannulated Holstein cows (550 ㎏ of mean body wt) were used for in situ trial, and a total of 96 nylon bags were retrieved from the rumen according to eight fermentation times (1, 3, 6, 9, 18, 24, 48 and 72 hr). The mean fermentation temperatures of TMRs by supplementation of anaerobic micoorganism co-cultures ranged from $22.97^{\circ}C$ to $26.07^{\circ}C$, and tended to increase steadily during the entire period. pH values of the F-TMRs ranged from 4.39 to 4.98 and tended to decrease with the extension of the fermentation period, and decreased by supplementation of synbiotics (p<0.05). The ammonia concentrations of F-TMRs were not affected by addition of synbiotic co-cultures during the early fermentation period (within 7 days), but was lowest (p<0.05) in T3 during the late fermentation periods (after 14 days). Lactic acid concentration of F-TMR was lowest in T3 at 1 day of fermentation, but was not different from treatments in the other fermentation days. Microbial growth rates of F-TMR reached a peak at 7 days of fermentation, and afterward tended to decrease. In in situ experiment, the DM disappearance rates were higher in T1 than the control during early fermentation times (within 3 hours), but was vice versa at 48 hours of fermentation (p<0.05). There was no significant difference in effective DM degradability among treatments. NDF and ADF disappearance rates in situ were similar to those of DM. From the above results, the supplementation of synbiotics, particularly the mould and bacteria synbiotics, resulted in improving the pH and concentration of lactic acid of F-TMR as parameters of fermentation compare to the control, and also had higher in situ disappearance rates of DM, NDF and ADF than the control at early fermentation time. However, effective DM degradability was not affected by supplementation of synbiotics.

• Journal of Korean Society of Environmental Engineers
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• v.38 no.6
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• pp.317-322
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• 2016

While PFOS sodium salt ($C_8F_{17}SO_3Na$) was not degraded by microorganisms for 28 days, the 4 alternatives were biodegraded at the rates of 21.6% for $C_{25}F_{17}H_{32}S_3O_{13}Na_3$, 20.5% for $C_{15}F_9H_{21}S_2O_8Na_2$, 15.8% for $C_{23}F_{18}H_{28}S_2O_8Na_2$ and 6.4% for $C_{17}F_9H_{25}S_2O_8Na_2$, respectively. The acute toxicity test using Daphnia magna was conducted for 48 hours, the half effective concentration ($EC_{50}$) of PFOS sodium salt ($C_8F_{17}SO_3Na$) was evaluated in 54.5 mg/L. While the 4 alternatives did not show any effect at 500.0 mg/L. The surface tension of the PFOS salt ($C_8F_{17}SO_3Na$) is 46.2 mN/m at a concentration of 500.0 mg/L. While the surface tension of the 4 alternatives was found to be superior to PFOS sodium salt ($C_8F_{17}SO_3Na$). The surface tension of $C_{23}F_{18}H_{28}S_2O_8Na_2$ (20.9 mN/m) has the lowest, followed by $C_{15}F_9H_{21}S_2O_8Na_2$ (23.4 mN/m), $C_{17}F_9H_{25}S_2O_8Na_2$ (27.3 mN/m), $C_{25}F_{17}H_{32}S_3O_{13}Na_3$ (28.2 mN/m). The four kinds of alternatives ($C_{15}F_9H_{21}S_2O_8Na_2$, $C_{17}F_9H_{25}S_2O_8Na_2$, $C_{23}F_{18}H_{28}S_2O_8Na_2$, $C_{25}F_{17}H_{32}S_3O_{13}Na_3$) were found to be superior to PFOS sodium salt ($C_8F_{17}SO_3Na$) in terms of biodegradation, Daphnia sp. acute toxicity and surface tension, and thus they were considered applicable as PFOS alternatives. Especially biodegradation rate of $C_{15}F_9H_{21}S_2O_8Na_2$, $C_{23}F_{18}H_{28}S_2O_8Na_2$ and $C_{25}F_{17}H_{32}S_3O_{13}Na_3$ was relatively high as 15.8~21.6%, and Daphnia sp. acute toxicity and surface tension were considerably superior (surface tension 39~55%) to PFOS sodium salt. Therefore, these alternatives are considered to be available as an alternative of PFOS.

• Korean journal of food and cookery science
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• v.30 no.6
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• pp.774-784
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• 2014

The purpose of this study was to present management guidelines for critical control points by analyzing microbiological hazardous elements through screening Potentially Hazardous Foods (PHF) menus in an effort improve the microbiological quality of foods prepared by restaurant operations. Steamed spinach with seasoning left at room temperature presents a range of risk temperatures which microorganisms could flourish, and it exceeded all microbiological safety limits in our study. On the other hand, steamed spinach with seasoning stored in a refrigerator had Aerobic Plate Counts of $2.86{\pm}0.5{\log}\;CFU/g$ and all other microbiological tests showed that their levels were below the limit. The standard plate counts of raw materials of lettuce and tomato were $4.66{\pm}0.4{\log}\;CFU/g$ and $3.08{\pm}0.4{\log}\;CFU/g$, respectively. Upon washing, the standard plate counts were $3.12{\pm}0.6{\log}\;CFU/g$ and $2.10{\pm}0.3{\log}\;CFU/g$, respectively, but upon washing after chlorination, those were $2.23{\pm}0.3{\log}\;CFU/g$ and $0.72{\pm}0.7{\log}\;CFU/g$, respectively. The standard plate counts of baby greens, radicchio and leek were $6.02{\pm}0.5{\log}\;CFU/g$, $5.76{\pm}0.1{\log}\;CFU/g$ and $6.83{\pm}0.5{\log}\;CFU/g$, respectively. After 5 minutes of chlorination, the standard plate counts were $4.10{\pm}0.6{\log}\;CFU/g$, $5.14{\pm}0.1{\log}\;CFU/g$ and $5.30{\pm}0.3{\log}\;CFU/g$, respectively. After 10 minutes of chlorination treatment, the standard plate counts were $2.58{\pm}0.3{\log}\;CFU/g$, $4.27{\pm}0.6{\log}\;CFU/g$, and $4.18{\pm}0.5{\log}\;CFU/g$, respectively. The microbial levels decreased as the time of chlorination increased. This study showed that the microbiological quality of foods was improved with the proper practices of time-temperature control, sanitization control, seasoning control, and personal and surface sanitization control. It also presents management guidelines for the control of potentially hazardous foods at the critical control points in the process of restaurant operations.

• Journal of Food Hygiene and Safety
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• v.26 no.4
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• pp.417-423
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• 2011

Salted Cabbage products purchased from different companies at 4 different districts in South Korea were detected in this study. Cabbage and salt are the main materials for kimchi manufacture. The results of general bacteria contaminated in the samples were $1.4{\times}10^5$, $6.4{\times}10^5$, $1.7{\times}10^7$, $3.6{\times}10^7$ CFU/g in cabbage and $2.7{\times}10^3$ CFU/g in salt, respectively. The results of coliforms were detected as $2.4{\times}10^4$ CFU/g, and there was no Escherichia coli in any sample. Staphylococcus aureus was detected in cabbage as $9.9{\times}10^2$, $8.0{\times}10^1$, and $3.0{\times}10^3$ CFU/g, Bacillus cereus was also found in cabbage as $4.1{\times}10^3$ and $1.0{\times}10^1$ CFU/g. The results of Campylobacter jejuni and Vibrio paraheamolyticus were $2.4{\times}10^6$ and $1.0{\times}10^4$ CFU/g in cabbage, respectively. $1.0{\times}10^3$ CFU/g for Yersinia enterocolitica was determined in salt. In case of Listeria monocytogenes, the results were $1.5{\times}10^1$, $1.1{\times}10^2$, and $4.5{\times}10^1$ CFU/g in cabbage. Total batcteria ranged from $1.4{\times}10^1$ to $4.4{\times}10^5$ CFU/g were detected in salting solution, from $1.5{\times}10^4$ to $1.2{\times}10^8$ CFU/g in dehydrated salted-cabbage, from $9.4{\times}10^4{\sim}1.3{\times}10^8$ CFU/g in minced salted-cabbage. The results of E. coli in samples from different companies were different from one to anther. The results of the contamination of S. aureus and B. cereus showed positive in salting solution and dehydrated salted-cabbage at a portion of companies. V. paraheamolyticus was detected in salting solution. The contamination of Y. enterocolitica ranged from $9.5{\times}10^2$ to $1.8{\times}10^3$ CFU/g in salting solution, from $1.7{\times}10^1$ to $2.7{\times}10^2$ CFU/g in dehydrated salted-cabbage, from $1.2{\times}10^2$ to $1.3{\times}10^8$ CFU/g in minced salted-cabbage. The contamination of L. monocytogenes ranged from $8.0{\times}10^2$ to $1.7{\times}10^4$ CFU/g in salting solution, from $2.8{\times}10^2$ to $1.2{\times}10^4$ CFU/g in dehydrated salted-cabbage. During the manufacture processing of Kim chi, microorganisms were detected in cabbages salted in different concentrations of salt solution at 8%, 10%, 12% and 15% for 5-20 hours. As the results, $3.5{\times}10^5-1.7{\times}10^6$, $3.4{\times}10^5-2.5{\times}10^6$, $5.4{\times}10^5-2.3{\times}10^6$, $4.0{\times}10^5-2.3{\times}10^6$ CFU/g were detected for E. coli in samples at different treatment conditions. $1.9{\times}10^4-4.1{\times}10^4$, $4.1{\times}10^3-2.8{\times}10^4$, $1.5{\times}10^3-7.8{\times}10^3$, $2.2{\times}10^4-6.6{\times}10^4$ CFU/g were detected for S. aureus in samples at different treatment conditions. Salmonella typhimurium was detected in salted cabbage with various salt concentration after salting for 5 hrs, the result ranged from $2.5{\times}10^5$ to $3.8{\times}10^6$ CFU/g, and change of microorganism was the smallest in salted cabbage under the concentration of salting solution at 10% for 15 hours. The cabbage salted in 10% salting solution for 15 hours were washed with water for 2 and 3 times, with chlorine for 3 times, and with acetic acid for 3 times. E. coli was detected in the samples washed with water for 2 and 3 times, washed with chlorine for 3 times. The contamination of S. aureus was $3.0{\times}10^5$ CFU/g in the samples washed with water for 2 times, $5.6{\times}10^3$ CFU/g in the samples washed with acetic acid for 3 times, $3.6{\times}10^5$ CFU/g in the samples washed with water for 3 times and same amount in the samples washed with chlorine for 3 times. According to the results, the contamination of S. aureus was $5.6{\times}10^3$ CFU/g lower in samples washed with chlorine and acetic acid than that in samples washed with water. In case of S. typhimurium, it has been detected in samples washed with water and chlorine, $3.0{\times}10^1$ CFU/g as the lowest concentration among all the samples was measured in the samples washed with acetic acid for 3 times.

• Applied Biological Chemistry
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• v.22 no.2
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• pp.65-90
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• 1979

This study was conducted to establish the brewing method which would be useful for the production of Kochuzang. Kojis, which were made from various materials and microorganisms under a covered condition, were investigated and compared. Yeasts (Saccharomyces rouxii and Torulopsis versatilis) were added to Kochuzang, and the enzyme activity, microflora, chemical composition, nitrogen content, alcohol content and free sugars of Kochuzang were investigated and analysed. The results obtained are as follows: 1. Koji making (1) Glutinous rice-soybean group was superior to glutinous rice group in the saccharogenic and liquefying amylase activities of three day-Koji. (2) Protease activity (acid, neutral and alkaline) of glutinous rice-soybean Koji, which was inoculated with Aspergillus oryzae A, was increased till the 5th day, while other groups showed maximum activity after the 3rd day. (3) The maximum cellulose activity of Aspergillus oryzae B-Koji and A-Koji was observed after the 2nd day and the 3rd day, respectively. High cellulose activity of Aspergillus oryzae B-Koji and A-Koji was respectively shown in glutinous rice group and glutinous rice-soybean group at maximum. (4) Compared with glutinous rice Koji, glutinous rice-soybean Koji gave larger number of yeast and aerobic bacteria. 2. Kochuzang Fermentation (1) Each Kochuzang group shoved different liquefying and saccharogenic amylase activities. The highest activities were generally shown in 10 to 40 days after mashing and remarkably reduced in the last stage of aging. (2) Protease activities of each group were strong in order of acid, neutral and alkaline protease. Especially acid protease showed highest activity at the 40th to 50th day Kochuzang. (3) Each group showed maximum cellulase activity in the 40th and 50th day-Kochuzang and then decreased. (4) Osmophilic yeast of yeast-added Kochuzang after one-month aging was distinctively outnumbered compared with non-yeast-added Kochuzang, but two groups were similar after two months. (5) Yeast-added group and non-added group gave almost the same number of halophilic lactic acid bacteria in Kochuzang, but the non-added group gave slightly larger number of aerobic bacteria than the yeast-added group. (6) Amino nitrogen contents in all test group were increased rapidly till the 60th day of Kochuzang aged. After that the contents were increased slowly. (7) Ethyl alcohol contents of 20day-fermented Kochuzang were high in order of Saccharomyces rouxii-added group, Torulopsis versatilis-added group, Saccharomyces rouxii and Torulopsis versatilis mixed group and non-yeast-added group. But all test group showed about 2% in ethyl alcohol content after 40days of aging. (8) Alcohol content in the 7 month-aged Kochuzang of all test groups was high in order of ethyl alcohol, n-butyl alcohol, n-propyl alcohol and iso-propyl alcohol. Torulopsis versatilis-added group had the highest value of ethyl alcohol, n-propyl alcohol and n-butyl alcohol. (9) Reducing sugar in Kochuzang was increased after 20 days of aging compared with the 10days-ferment. The reducing sugar content in Saccharomyces rouxii-added group was distinctively small compared with that of other groups, decreasing after 30days of aging. (10) Rhamnose, fructose, glucose and maltose were isolated from the 10 day fermented Kochuzang. Raffinose was also found after 300 days-aged group, and fructose content was high in the 300days-aged Kochuzang. However, glucose content was smaller than that of 10days-fermented Kochuzang. (11) For the organoleptic tests of Kochuzang, taste, flavour and color of yeast-added group were superior to the non yeast-added group. Especially the complex yeast group among the yeast added groups were the best of all. Yeast-added group after 300 days of aging took higher paint in flavour test than that of non-added group. Therefore, brewing method like complex yeast added group seems to be advantageous for short time brewing Kochuzang.

• 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 $60^{\circ}C$.

• Applied Biological Chemistry
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• v.10
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• pp.69-100
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• 1968

1. In order to investigate on the microflora and enzyme activity of mold wheat 'Nuruk' , the major source of microorganisms for the brewing of Takju (a Korean Sake), two samples of Nuruk, one prepared at the College of Agriculture, Chung Nam University (S) and the other perchased at a market (T), were taken for the study. The molds, aerobic bacteria, lactic acid bacteria, and yeasts were examined and counted. The yeasts were classified by the treatment with TTC (2, 3, 5 triphenyltetrazolium chloride) agar that yields a varied shade of color. The amylase and protease activities of Nuruk were measured. The results were as the followings. a) In the Nuruk S found were: Aspergillus oryzae group, $204{\times}10^5$; Black Aspergilli, $163{\times}10^5$; Rhizogus, $20{\times}10^5$; Penicillia, $134{\times}10^5$; Areobic bacteria, $9{\times}10^6-2{\times}10^7$; Lactic acid bacteria, $3{\times}10^4$ In the Nuruk T found were: Aspergillus oryzae group, $836{\times}10^5$; Black Aspergilli, $286{\times}10^5$; Rhizopus, $623{\times}10^5$; Penicillia, $264{\times}10^5$; Aerobic bacteria, $5{\times}10^6-9{\times}10^6$; Lactic acid bacteria, $3{\times}10^4$ b) Eighty to ninety percent of the aerobic bacteria in Nuruk S appeared to belong to Bacillus subtilis while about 70% of those in Nuruk T seemed to be spherical bacteria. In both Nuruks about 80% of lactic acid bacteria were observed as spherical ones. c) The population of yeasts in 1g. of Nuruk S was about $6{\times}10^5$, 56.5% of which were TTC pink yeasts, 16% of which were TTC red pink yeasts, 8% of which were TTC red yeasts, 19.5% of which were TTC white yeasts. In Nuruk T(1g) the number of yeasts accounted for $14{\times}10^4$ and constituted of 42% TTC pink. 21% TTC red pink 28% TTC red and 9% TTC white. d) The enzyme activity of 1g Nuruk S was: Liquefying type Amylase, $D^{40}/_{30},=256$ W.V. Saccharifying type Amylase, 43.32 A.U. Acid protease, 181 C.F.U. Alkaline protease, 240C.F.U. The enzyme activity of 1g Nuruk T was: Liquefying type Amylase $D^{40}/_{30},=32$ W.V. Saccharifying type amylase $^{30}34.92$ A.U. Acid protease, 138 C.F.U. Alkaline protease 31 C.F.U. 2. During the fermentation of 'Takju' employing the Nuruks S and T the microflora and enzyme activity throughout the brewing were observed in 12 hour intervals. TTC pink and red yeasts considered to be the major yeasts were isolated and cultured. The strains ($1{\times}10^6/ml$) were added to the mashes S and T in which pH was adjusted to 4.2 and the change of microflora was examined during the fermentation. The results were: a) The molds disappeared from each sample plot since 2 to 3 days after mashing while the population of aerobic bacteria was found to be $10{\times}10^7-35{\times}10^7/ml$ inS plots and $8.2{\times}10^7-12{\times}10^7$ in plots. Among them the coccus propagated substantially until some 30 hours elasped in the S and T plots treated with lactic acid but decreased abruptly thereafter. In the plots of SP. SR. TP. and TR the coccus had not appeared from the beginning while the bacillus showed up and down changes in number and diminished by 1/5-1/10 the original at the end stage. b) The lactic acid bacteria observed in the S plot were about $7.4{\times}10^7$ in number per ml of the mash in 24 hours and increased up to around $2{\times}10^8$ until 3-4 days since. After this period the population decreased rapidly and reached about $4{\times}10^5$ at the end, In the plot T the lactic acid becteria found were about $3{\times}10^8$ at the period of 24 fours, about $3{\times}10$ in 3 days and about $2{\times}10^5$ at the end in number. In the plots SP. SR. TP, and TR the lactic acid bacteria observed were as less as $4{\times}10^5$ at the stage of 24 hours and after this period the organisms either remained unchanged in population or ceased to exist. c) The maiority of lactic acid bacteria found in each mash were spherical and the change in number displayed a tendency in accordance with the amount of lactic acid and alcohol produced in the mash. d) The yeasts had showed a marked propagation since the period of 24 hours when the number was about $2{\times}10^8$ ㎖ mash in the plot S. $4{\times}10^8$ in 48 hours and $5-7{\times}10^8$ in the end period were observed. In the plot T the number was $4{\times}10^8$ in 24 hours and thereafter changed up and down maintaining $2-5{\times}10^8$ in the range. e) Over 90% of the yeasts found in the mashes of S and T plots were TTC pink type while both TTC red pink and TTC red types held range of $2{\times}10-3{\times}10^7$ throughout the entire fermentation. f) The population of TTC pink yeasts in the plot SP was as $5{\times}10^8$ much as that is, twice of that of S plot at the period of 24 hours. The predominance in number continued until the middle and later stages but the order of number became about the same at the end. g) Total number of the yeasts observed in the plot SR showed little difference from that of the plot SP. The TTC red yeasts added appeared considerably in the early stage but days after the change in number was about the same as that of the plot S. In the plot TR the population of TTC red yeasts was predominant over the T plot in the early stage which there was no difference between two plots there after. For this reason even in the plot w hers TTC red yeasts were added TTC pink yeasts were predominant. TTC red yeasts observed in the present experiment showed continuing growth until the later stage but the rate was low. h) In the plot TP TTC pink yeasts were found to be about $5{\times}10^8$ in number at the period of 2 days and inclined to decrease thereafter. Compared with the plot T the number of TTC pink yeasts in the plot TP was predominant until the middle stage but became at the later stage. i) The productivity of alcohol in the mash was measured. The plot where TTC pink yeasts were added showed somewhat better yield in the earely stage but at and after the middle stage the difference between the yeast-added and the intact mashes was not recognizable. And the production of alcohol was not proportional to the total number of yeasts present. j) Activity of the liquefying amylase was the highest until 12 hours after mashing, somewhat lowered once after that, and again increased around 36-48 hours after mashing. Then the activity had decreased continuously. Activity of saccharifying amylase also decreased at the period of 24 hours and then increased until 48 hours when it reached the maximum. Since, the activity had gradually decreased until 72 hours and rapidly so did thereafter. k) Activity of alkaline protease during the fermentation of mash showed a tendency to decrease continusously although somewhat irregular. Activity of acid protease increased until hours at the maximum, then decreased rapidly, and again increased, the vigor of acid protease showed better shape than that of alkaline protease throughout. 3. TTC pink yeasts that were predominant in number, two strains of TTC red pink yeasts that appeared throughout the brewing, and TTC red yeasts were identified and the physiological characters examined. The results were as described below. a) TTC pinkyeasts (B-50P) and two strains of TTC red pink yeasts (B-54 RP & B-60 RP) w ere identified as the type of Saccharomyces cerevisiae and TTC pink red yeasts CB-53 R) were as the type of Hansenula subpelliculosa. b) The fermentability of four strains above mentioned were measured as follows. Two strains of TTC red pink yeasts were the highest, TTC pink yeasts were the lowest in the fermantability. The former three strains were active in the early stage of fermentation and found to be suitable for manufacturing 'Takju' TTC red yeasts were found to play an important role in Takju brewing due to its strong ability to produce esters although its fermentability was low. c) The tolerance against nitrous acid of strains of yeast was marked. That against lactic acid was only 3% in Koji extract, and TTC red yeasts showed somewhat stronger resistance. The tolerance against alcohol of TTC pink and red pink yeasts in the Hayduck solution was 7% while that in the malt extract was 13%. However, that of TTC red yeasts was much weaker than others. Liguefying activity of gelatin by those four strains of yeast was not recognized even in 40 days. 4. Fermentability during Takju brewing was shown in the first two days as much as 70-80% of total fermentation and around 90% of fermentation proceeded in 3-4 days. The main fermentation appeared to be completed during :his period. Productivity of alcohol during Takju brewing was found to be apporximately 65% of the total amount of starch put in mashing. 5. The reason that Saccharomyces coreanuss found be Saito in the mash of Takju was not detected in the present experiment is considered due to the facts that Aspergillus oryzae has been inoculated in the mold wheat (Nuruk) since around 1930 and also that Koji has been used in Takju brewing, consequently causing they complete change in microflora in the Takju brewing. This consideration will be supported by the fact that the original flavor and taste have now been remarkably changed.