• Title/Summary/Keyword: Bacillus

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Isolation and Characterization of Lactic acid bacteria Leuconostoc mesenteroides DB3 from Camellia japonica Flower (백꽃으로부터 분리한 Leuconostoc mesenteroides DB3의 특성)

  • Sam Woong Kim;Da Hye Shin;Sang Wan Gal;Kyu Ho Bang;Da Som Kim;Won-Jae Chi
    • Journal of Life Science
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    • v.33 no.11
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    • pp.915-922
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    • 2023
  • Lactic acid bacteria (LAB) are widespread in a variety of environments including fermented dairy products, gastroinstetinal and urogenital tracts of human and animals, plant, soil and water. Leuconostoc mesenteroides DB3 was detected by the strongest antibacterial activities among 24 Leuconostoc strains isolated from Camellia japonica flowers. Acid tolerance of L. mesenteroides DB3 existed up to pH 2.5, but the resistance did not show at pH 2.0, which relatively excellent acid resistance existed. Bile acid tolerance was very stable within the test range to 1.2%. L. mesenteroides DB3 exhibited the optimal growth at 30℃, and showed a slight slow growth when compared with L. mesenteroides KCTC3505, which reached a stationary phase at 18 hr. The pH was changed along with the growth curve, but was maintained above pH 3.98. L. mesenteroides DB3 had higher initial antibacterial activities when compared to L. mesenteroides KCTC3505, but it showed similar activities with the standard strain after the latter part of the logarithmic growth phase. Although lactic acid production in L. mesenteroides DB3 was induced by lower amount in the initial part to the standard strain, it was exhibited by similar amounts after the late logarithmic growth phase. Muicin adhesion of L. mesenteroides DB-3 maintained superior to L. mesenteroides KCTC3505. Both strains showed excellent emulsification ability for kerosene. In summary, we evaluate that L. mesenteroides DB-3 has a high potential for application as probiotics owing to its excellent antibacterial activity, acid resistance, bile acid resistance, and muicin adhesion.

Effects of Microbial Fermentation on the Antioxidant Activities of Protaetia brevitarsis Larvae (미생물 발효가 흰점박이꽃무지(Protaetia brevitarsis) 유충의 항산화 활성에 미치는 영향)

  • Han Bi Kim;Hye Soo Kim;Soo Jeong Cho
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1052-1061
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    • 2023
  • This study was carried out to evaluate the effect of fermentation by B. subtilis (BPLE), L. brevis (LPLE), S. cerevisiae (SPLE) and C. militaris (CPLE) on the antioxidant activity of Protaetia brevitarsis larvae fed with mushroom substrates (king oyster mushroom). The total polyphenol content of Protaetia brevitarsis larvae (PLE), BPLE, LPLE, SPLE and CPLE were 58.07±0.67, 83.33±0.98, 79.21±1.32, 61.02±0.87 and 57.90±1.02 mg GAEs/extract g, respectively. The flavonoid contents of the PLE, BPLE, LPLE, SPLE and CPLE were 17.35±1.57, 19.49±0.95, 16.90±1.57, 18.12±0.95 and 16.99±0.95 mg QEs/extract g, respectively. The DPPH radical scavenging activity showed no significant difference between the PLE, BPLE, LPLE, SPLE and CPLE at a concentration of 0.2 mg/ml. However, at a concentration of 0.4 mg/ml or more, the DPPH radical scavenging activity of the BPLE and LPLE was higher than that of the PLE. The reducing power of the BPLE and LPLE was also higher than that of the PLE, and more than twice as high at a concentration of 0.8 mg/ml or more. The ORAC value of the BPLE (79.77±0.82 uM TEs/extract g) was higher than that of the PLE (61.34±0.97 uM TEs/extract g). A WST-1 assay of the RAW 264.7 cells indicated that the PLE, BPLE, LPLE, SPLE and CPLE showed no cytotoxicity.

Characteristics of Leuconostoc spp. isolated from radish kimchi and its immune enhancement effect (무김치에서 분리한 Leuconostoc 속의 특성과 면역증강 효과)

  • Seoyeon Kwak;Seongeui Yoo;Jieon Park;Woosoo Jeong;Hee-Min Gwon;Soo-Hwan Yeo;So-Young Kim
    • Food Science and Preservation
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    • v.30 no.6
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    • pp.1082-1094
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    • 2023
  • The purpose of this study was to examine the characteristics of Leuconostoc spp. isolated from radish kimchi and to investigate the potential for the use of functional ingredients by evaluating enzymatic characteristics, safety, and immune-enhancing effects among the isolates, including Lactobacillus rhamnosus ATCC53103 (LGG) as a control strain. All test strains exhibited β-glucosidase enzyme activity that releases β-1,4 sugar chain bonds. In addition, as a result of antibiotic resistance assay among the isolates, MIC values on 8 antibiotics were below compared to the EFSA standard, and hemolytic experiments confirmed that all showed gamma hemolysis without hemolytic ability. As a result of the antibacterial activity experiment, the Leu. mesenteroides K2-4 strain showed a higher activity than LGG against Bacillus cereus and Staphylococcus aureus. Additionally, the activity of the NF-kB/AP-1 transcription factor increased when the isolates were treated in macrophage RAW cells. These results were related to increasing the high mRNA expression levels on TNF-α and IL-6 by Leu. mesenteroides K2-4 strain to be treated at low concentration. Consequently, we suggest that it will be useful as a candidate for functional food ingredients.

Microbiological and Enzymological Studies on Takju Brewing (탁주(濁酒) 양조(釀造)에 관(關)한 미생물학적(微生物學的) 및 효소학적(酵素學的) 연구(硏究))

  • Kim, Chan-Jo
    • 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.

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Seasonal Variations of Water Quality in the Lower Part of the Nagdong River (낙동강 하류수질의 계절적 변화)

  • KIM Yong-Gwan;SHIM Hye-Kung;CHO Hak-Rae;YOU Sun-Jae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.6
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    • pp.511-522
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    • 1984
  • The Nagdong is one of the biggest rivers in Korea, which is very important water source not only for tap water of Pusan city but also for the industrial water. Therefore, authors tried to check the water quality year by year. In this experiment one hundred and twenty water samples collected from August 1983 to July 1984 were analyzed bacteriologically and physiologically. Fifteen sampling stations were established between near Samrangjin and estuary of the river. To evaluate the water quality, temperature, pH, chloride ion, salinity, chemical oxygen demand (COD), electrical conductivity, nutrients, total coliform, fecal coliform, fecal streptococcus, viable cell count and bacterial flora were observed. The variation of water temperature was ranged $-1.5{\sim}29.0^{\circ}C$ (Mean value $13.9{\sim}16.5^{\circ}C$), it in spring was higher as $10{\sim}15^{\circ}C$ about $10^{\circ}C$ than in winter and it in autumm was very stabilized as about $20^{\circ}C$ at each station. The pH variation of the samples was ranged $6.68{\sim}9.15$. The range of concentration of chloride ion and salinity varied $7.4{\sim}l,020.5$ mg/l and $1.05{\sim}33.0\%0$, respectively. Especially, salinity of the 3rd water war was the higher than others as $25.76{\sim}31.58\%0$. COD was ranged $1.45{\sim}14.94$ mg/l and the lower part of the Nagdong River was heavily contaminated by domesitc sewage and waste water from the adjacent factor area. The range of electrical conductivity was $1.360{\times}10^2{\sim}5.650{\times}10^4{\mu}{\mho}/cm$ and that was by far higher the estuary than the upper. Concentration of nutrients were $0.008{\sim}0.040$ mg/l (Mean value $0.019{\sim}0.068$ mg/l) for $NO_2-N,\;0.038{\sim}5.253$ mg/l ($0.351{\sim}2.347$ mg/l) for $NO_3-N,\;0.100{\sim}2.685$ mg/l($0.117{\sim}1.380$ mg/l) for $NH_4-N,\;0.003{\sim}0.084$ mg/l($0.014{\sim}0.065$ mg/l) for $PO_4-P$ and $0.154{\sim}6.123$ mg/l ($1.165{\sim}3.972$ mg/l) for $SiO_2-Si$, respectively. Usually nutrients contents of the water in the upper part(included station 1 to 5) were higher than those of the estuarine area. The bacterial density of the samples ranged 7.3 to 460,000/100 ml for total coliforms, 3.6 to 460,000/100 ml for fecal coliform, $0{\sim}46,000/100ml$ for fecal streptococcus and $<30{\sim}1.2{\times}10^5/ml$ for viable cell count. Composition of coliform was $28\%$ Escherichia coli group, $18\%$ Citrobacter freundii group, $31\%$ Enterobacter aerogenes group and $22\%$ others. Predominant species among the 659 strains isolated from the samples were Pseudomonas spp. ($42\%$), Flavobacterium spp. ($20\%$) and Moraxella spp. ($12\%$).

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Change of Harmful Micnoorganisms in Pickling Process of Salted Cabbage According to Salting and Washing Conditions (배추김치의 절임공정 조건에 따른 위해미생물 변화)

  • Kim, Jin-Hee;Lee, Yu-Keun;Yang, Ji-Young
    • 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.

Implementation of integrated monitoring system for trace and path prediction of infectious disease (전염병의 경로 추적 및 예측을 위한 통합 정보 시스템 구현)

  • Kim, Eungyeong;Lee, Seok;Byun, Young Tae;Lee, Hyuk-Jae;Lee, Taikjin
    • Journal of Internet Computing and Services
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    • v.14 no.5
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    • pp.69-76
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    • 2013
  • The incidence of globally infectious and pathogenic diseases such as H1N1 (swine flu) and Avian Influenza (AI) has recently increased. An infectious disease is a pathogen-caused disease, which can be passed from the infected person to the susceptible host. Pathogens of infectious diseases, which are bacillus, spirochaeta, rickettsia, virus, fungus, and parasite, etc., cause various symptoms such as respiratory disease, gastrointestinal disease, liver disease, and acute febrile illness. They can be spread through various means such as food, water, insect, breathing and contact with other persons. Recently, most countries around the world use a mathematical model to predict and prepare for the spread of infectious diseases. In a modern society, however, infectious diseases are spread in a fast and complicated manner because of rapid development of transportation (both ground and underground). Therefore, we do not have enough time to predict the fast spreading and complicated infectious diseases. Therefore, new system, which can prevent the spread of infectious diseases by predicting its pathway, needs to be developed. In this study, to solve this kind of problem, an integrated monitoring system, which can track and predict the pathway of infectious diseases for its realtime monitoring and control, is developed. This system is implemented based on the conventional mathematical model called by 'Susceptible-Infectious-Recovered (SIR) Model.' The proposed model has characteristics that both inter- and intra-city modes of transportation to express interpersonal contact (i.e., migration flow) are considered. They include the means of transportation such as bus, train, car and airplane. Also, modified real data according to the geographical characteristics of Korea are employed to reflect realistic circumstances of possible disease spreading in Korea. We can predict where and when vaccination needs to be performed by parameters control in this model. The simulation includes several assumptions and scenarios. Using the data of Statistics Korea, five major cities, which are assumed to have the most population migration have been chosen; Seoul, Incheon (Incheon International Airport), Gangneung, Pyeongchang and Wonju. It was assumed that the cities were connected in one network, and infectious disease was spread through denoted transportation methods only. In terms of traffic volume, daily traffic volume was obtained from Korean Statistical Information Service (KOSIS). In addition, the population of each city was acquired from Statistics Korea. Moreover, data on H1N1 (swine flu) were provided by Korea Centers for Disease Control and Prevention, and air transport statistics were obtained from Aeronautical Information Portal System. As mentioned above, daily traffic volume, population statistics, H1N1 (swine flu) and air transport statistics data have been adjusted in consideration of the current conditions in Korea and several realistic assumptions and scenarios. Three scenarios (occurrence of H1N1 in Incheon International Airport, not-vaccinated in all cities and vaccinated in Seoul and Pyeongchang respectively) were simulated, and the number of days taken for the number of the infected to reach its peak and proportion of Infectious (I) were compared. According to the simulation, the number of days was the fastest in Seoul with 37 days and the slowest in Pyeongchang with 43 days when vaccination was not considered. In terms of the proportion of I, Seoul was the highest while Pyeongchang was the lowest. When they were vaccinated in Seoul, the number of days taken for the number of the infected to reach at its peak was the fastest in Seoul with 37 days and the slowest in Pyeongchang with 43 days. In terms of the proportion of I, Gangneung was the highest while Pyeongchang was the lowest. When they were vaccinated in Pyeongchang, the number of days was the fastest in Seoul with 37 days and the slowest in Pyeongchang with 43 days. In terms of the proportion of I, Gangneung was the highest while Pyeongchang was the lowest. Based on the results above, it has been confirmed that H1N1, upon the first occurrence, is proportionally spread by the traffic volume in each city. Because the infection pathway is different by the traffic volume in each city, therefore, it is possible to come up with a preventive measurement against infectious disease by tracking and predicting its pathway through the analysis of traffic volume.