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Activity and Survival of the Natural Bacteria under the Stressed Conditions Detected by Bioluminescent Phenotype (스트레스 하의 자연세균의 활성 및 생존의 발광표현형을 이용한 탐지)

  • Park, Kyoung-Je;Yoon, Hye-Young;Chun, Se-Jin;Lee, Ho-Sa;Lee, Dong-Hun;Jahng, Deokjin;Lee, Kyu-Ho
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.154-161
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    • 1998
  • To investigate whether the introduced genetic marker is useful to detect the survivalship and activity of the natural bacteria under the stressed conditions, one Gram-negative isolate, KP964 was transformed to the luminous phenotype by transferring luxAB gene. Under the starvation-stress this luminous bacterial culturability (determined by colony-forming-units [CFU] on agar plate) decreased rapidly below the detection limit by 37 days, while its total cell number (determined by AODC) remained almost the same as its initial inocular size. At that time period, the viable cell number was estimated to be 1400 times higher than its CFU number. The bioiuminescence (determined by relative light units [RLU]) produced under the same condition was also monitored and found to decrease more rapidly than the culturability by 5-fold. Under the other stresses, e.g., osmotic shocks, acid shock, and exposure to toxic chemicals, this bacterial strain did not show the reliable correlation between CFU and RLU. These results might not suggest the direct estimation of bioiuminescence from the stressed bacteria be an index of both the survivalship and its activity. However, when the stressed bacterial cells were incubated under the favorable condition by relieving from the existing stress, the potential bioiuminescence (the lag periods before the increase of bioiuminescence, the increase rates of bioiuminescence, and the maximal levels of bioiuminescence) was shown to be highly dependent upon the strengths of the stresses exposed to the bacterial cells. Therefore, analysis of the potential bioiuminescence from the stressed bacteria revealed good relationships with survival as well as activity.

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Distribution and Favorable Conditions for Mycelial Growth of Cordyceps pruinosa in Korea (한국에 자생하는 붉은자루동충하초(Cordyceps pruinosa)의 분포와 균사생장에 적합한 조건)

  • Shin, Jae-Chul;Shrestha, Bhushan;Lee, Won-Ho;Park, Young-Jin;Kim, Soo-Young;Jeong, Gwang-Ryel;Kim, Ho-Kyung;Kim, Tae-Woong;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.79-88
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    • 2004
  • Cordyceps pruinosa grows upon dead pupae of Lepidoptera and produces one or $3{\sim}4$ club-shaped stromata per host. The stromata have distinct club-shaped head and long stalk. The length of stromata varies from $1{\sim}3\;cm$. Apical head consists of densely crowded semi-immersed perithecia, which are $360{\sim}400\;{\times}\;180{\sim}200\;{\mu}m$ in size. Asci are $150\;{\mu}m$ in length and $2.8{\sim}3\;{\mu}m$ in diameter. Ascospores, which are $124{\sim}141\;{\mu}m$ in length, have thin thread-like structures in the middle with part-spores attached on both sides. Each ascospore does not separate into part-spores after dispersal, but each part-spore germinates and together develops a colony. The imperfect form produces phialides of $15{\sim}24\;{\times}\;2{\sim}3\;{\mu}m$ size, with spherical or spindle shaped conidia of $4{\sim}6\;{\times}\;1.8{\sim}2.4\;{\mu}m$ size, The anamorph was identified as Mariannaea elegans Samson. YMA and SDAY agar media with pH 7 was produced abundant mycelial growth with high density. Best mycelial growth was observed when dextrin was used as a carbon source. Lactose, saccharose and sucrose also produced high mycelial growth. Peptone, yeast extract and tryptone produced abundant mycelial growth, when used as nitrogen sources. Highest mycelial growth and density was observed when C/N ratio was 1 : 1 at the concentration of 12.5 g/l each. $KH_2PO_4$ was the best mineral source for mycelial growth. Highest mycelial dry wt. was produced in YM and SDAY broths. Optimum inoculum for 100 ml of liquid broth was 6 mycelial discs. Similarly, optimum liquid culture period was 7 days.

Fungicide Resistance of Fusarium fujikuroi Isolates Isolated In Korea (우리나라에서 분리한 벼 키다리병균(Fusarium fujikuroi)의 살균제 저항성)

  • Lee, Yong-Hwan;Kim, So-Yeon;Choi, Hyo-Won;Lee, Myeong-Ji;Ra, Dong-Soo;Kim, In-Seon;Park, Jin-Woo;Lee, Se-Weon
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.427-432
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    • 2010
  • Rice bakanae disease caused by Fusarium fujikuroi is one of the most serious rice diseases in Korea. From 2006 to 2009, 118 F. fujikuroi isolates were collected from various regions of rice fields in Korea. Resistance assay of 118 F. fujikuroi isolates to prochloraz, tebuconazole, and benomyl, were performed using agar dilution method. To investigate inhibitory effects of the fungicides, minimum inhibitory concentration of mycelial growth (MIC) and effective concentration inhibiting mycelial growth by 50% ($EC_{50}$) for 118 isolates were calculated using Sigmaplot 8.02 (Antro, SPSS UK, Ltd). Based on the means of $EC_{50}$ values, baseline resistance values were determined as $0.5{\mu}g{\cdot}mL^{-1}$ for prochloraz, $5.0{\mu}g{\cdot}mL^{-1}$ for tebuconazole and $2.5{\mu}g{\cdot}mL^{-1}$ for benomyl. Number of resistant isolates to each fungicide was 17, 19 and 43 for prochloraz, tebuconazole and benomyl, respectively. Furthermore, 4 isolates showed the double resistance to both prochloraz and tebuconazole, 6 isolates to prochloraz and benomyl, and 11 isolates to tebuconazole and benomyl. Isolates CF366 and LF335 isolated from Gyeongbuk province were resistant to the three fungicides tested, prochloraz, tebuconazole and benomyl.

Microbial and Pathogenic Contamination of Ready-to-eat Fresh Vegetables in Korea (한국에 유통중인 신선편이 채소류의 미생물 품질 및 병원성 세균의 오염도 조사)

  • Bae, Young-Min;Hong, Yu-Jin;Kang, Dong-Hyun;Heu, Sung-Gi;Lee, Sun-Young
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.161-168
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    • 2011
  • The purpose of this study was to evaluate microbiological contamination of fresh vegetables in Korea. Twenty types of vegetables were tested for total aerobic bacteria, coliforms, Escherichia coli, yeast and mold, and pathogenic bacteria such as Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Salmonella, E. coli O157:H7, Cronobacter sakazakii, Shigella, and Campylobacter. Levels of total aerobic bacteria and coliform on 20 vegetables were between 3.74 and 8.04 log CFU/g, and 0.16 and 5.02 log CFU/g, respectively. The highest contamination levels of total aerobic bacteria were observed on water dropwort, sprouts, mungbean sprout, and ballflower root. There was no significant difference (p>0.05) in microbial contamination levels of total aerobic count, coliform, E. coli, yeast and mold between organic and nonorganic vegetables. When isolation methods using selective agars were applied, L. monocytogenes, B. cereus, Salmonella and Campylobacter were isolated from some fresh vegetable samples. Results of API kit tests showed that L. monocytogenes was identified on Chinese cabbage, cucumber, soybean sprouts, and iceberg lettuce while Salmonella was identified on Korean leek. Furthermore, Campylobacter jejuni was also identified in more than 50 of the 100 samples. However, when positive samples from API kit were tested for real-time PCR or 16S rRNA sequencing method, only B. cereus from perilla leaf, carrot, water dropwort, and sprouts showed positive results. These results indicate that selective agar and API kit detection methods might result in false positive results for some pathogens. Therefore, studies need to improve isolation or confirmation methods for such pathogens.

Characteristics and Survival of Genus Vibrio Isolated in the Intertidal Zone of the Yellow Sea near Kunsan (군산인근해역에서 분리동정된 Vibrio 속의 특성과 해수에서의 생존)

  • 왕혜영;이건형
    • Korean Journal of Environmental Biology
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    • v.17 no.4
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    • pp.439-448
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    • 1999
  • To investigate the population dynamics and survival of Genus Vibrio, population densities of aerobic saprophytic bacteria and Vibrio groups were measured 4 times in the intertidal waters of the Yellow Sea near Kunsan from November, 1997 to June, 1998. The distribution of heterotrophic bacteria during the survey periods by plate count and direct count method ranged from 1.2$\pm$0.6$\times$10$^3$~2.0$\pm$1.5$\times$10$^4$CFU ml­$^1$and from 6.0$\pm$4.0$\times$10$^{5}$ ~1.9$\pm$1.5$\times$10$^{7}$ cells ml­$^1$, respectively. Vibrio groups were distributed in the range of 1$\times$10 and 6$\pm$2.2$\times$10$^2$CFU ml­$^1$. The proportion of Vibrio groups to total heterotrophic bacteria was between 0.1 and 6% during the survey periods. A total of 51 isolates was obtained from TCBS agar plates and identified to species level by Biolog Identification System$^{TM}$. As a result, dominant genera were V, mediterranei, V aitguillarum, tr metschnikovii, and V. parahaemolyticus, and isolates were clustered into 26 groups based on the relatedness of average linkage clustering method at 70% level. As for the susceptibility of 51 isolates to 7 kinds of antibacterial agents (gentamicin, ampicillin, chlorarnphenicol, streptomycin, kanamycin, tetracycline, carbenicillin), 96% of isolates showed high resistance to more than one antibiotics and 65% of isolates contained a plasmid, of which size was observed greater than 12 kb, The number of cells of 3 tested strains (V. anguillarum, V. vulnificus, and V. metschnikovii) in filtered aged seawater decreased by approximately 1 to 5 orders of magnitude during 30-d incubation. In most cases, the numbers of cells decreased rapidly until day 3, then decreased slowly by day 30. The number of cells incubated at 15$^{\circ}C$ showed higher survival than those at 4$^{\circ}C$ and $25^{\circ}C$. These results may be considered for the basic supporting data in the risk assessment of vibriosis in summer.r.

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Two Strains of Colletotrichum gloeosporioides Penz. Causing Anthracnose on Pepper Fruits (고추탄저병균 Colletotrichum gloeosporioides Penz.의 2계통)

  • Kim Wan Gyu;Cho Eui Kyoo;Lee Eun Jong
    • Korean Journal Plant Pathology
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    • v.2 no.2
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    • pp.107-113
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    • 1986
  • Each of 48 monoconidial isolates of Colletotrichum gloeosporioides Penz. obtained from diseased fruits of pepper was classified into strain G or strain R based upon pathogenicity to green and red fruits, morphology of conidia, and cultural characteristics in potato dextrose agar. The strain G was designated for isolates to cause anthracnose symptoms both on green and red fruits. All isolates of the strain G produced conidia abundantly. but produced no perithecia and setae in PDA. Conidia of all isolates in the strain G were attenuated or round at one end. The optimum temperature for mycelial growth of strain G was $26-28^{\circ}C$. The mycelia of strain G in PDA appeared to be whitish when young, and turned to be dark in old culture. Symptoms on pepper fruits caused by the strain G were somewhat sunken to be circular to elliptical lesions. Yellowish conidial masse were observed at the center of lesions, and the lesions turned to irregular shape and to reddish brown color in the later stage of disease development. No setae were visible on the acervuli. The strain R was designated for isolates to cause anthracnose symptoms only on red fruits of pepper. All isolates of the strain R produce conidia, and perithecia of Glomerella cingulata (Stonem.) Spauld. & v. Sch. in PDA. Some isolates of the strain R produced setae in culture under fluorescent light. Conidia of all isolates in the strain R were round and blunt at the ends. The optimum temperature for mycelial growth of strain R was the same as that of strain G. The mycelial growth of strain R was faster than that of strain G in PDA. The mycelia of strain R in PDA appeared to be gray to dark. Symptoms on pepper fruits caused by the strain R were circular to irregular black ring-spots Short setae or no setae were visible on the acervuli.

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Isolation and Characterization of Starch-hydrolyzing Pseudoalteromonas sp. A-3 from the Coastal Sea Water of Daecheon, Republic of Korea (대한민국 대천 해안에서 분리한 전분 분해능을 갖는 Pseudoalteromonas sp. A-3 균주의 특징 및 동정)

  • Chi, Won-Jae;Park, Da-Yeon;Jeong, Sung-Cheol;Chang, Yong-Keun;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.317-323
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    • 2011
  • Strain A-3, an amylase-producing bacteria, was isolated from coastal seawater near Daecheon in the Republic of Korea. It was seen to possess a single polar flagella and grow well, on ASW-YP agar plates, at temperatures of between $20-37^{\circ}C$. However, it grew more slowly at the temperatures of $15^{\circ}C$ and $40^{\circ}C$. Similarly, it was observed to grow abundantly, in an Artificial Sea Water-Yeast extract-Peptone (ASW-YP) liquid medium, in a pH range of 6-9, but not grow at pHs of 4-5 and a pH of 10. Strain A-3 was noted as being close to Pseudoalteromonas phenolica O-$BC30^T$, Pseudoalteromonas luteoviolacea $NCIMB1893^T$, Pseudoalteromonas rubra $ATCC29570^T$, and Pseudoalteromonas byunsanensis $FR1199^T$, with 98.30%, 97.86%, 97.78%, and 97.25% similarities respectively, in its 16S rRNA sequence. A phylogenetic tree revealed that strain A-3 and P. phenolica O-$BC30^T$ belong to a clade. However, strain A-3 differed from P. phenolica O-$BC30^T$ in relation to a number of physiological characteristics. Strain A-3 exhibited no growth above 5% NaCl concentrations, no utilization of D-glucose, D-mannose, D-maltose, or D-melibose, and no lipase (C-14) activity. All of these properties strongly indicate that strain A-3 is distant from P. phenolica O-$BC30^T$ and thus led us to name it Pseudoalteromonas sp. A-3. Pseudoalteromonas sp. A-3 produces ${\alpha}$-amylase throughout growth. Maximal amylase activities of 144.48 U/mL and 149.20 U/mL were seen at pH 7.0 and $37^{\circ}C$, respectively. Pseudoalteromonas sp. A-3's high, stable production of ${\alpha}$-amylase in addition to its biochemical features, such as alkalitolerance, suggest that it is a good candidate for industrial applications.

Studies on Production of Heteropolysaccharide by Mutant of Xanthomonas malvacearum (Xanthomonas malvacearum 돌연변이주(突然變異株)의 Heteropolysaccharide 생산성(生産性)에 관(關)하여)

  • Lee, Ke-Ho;Kim, Mi-Sun;Park, Chan-Yung
    • Applied Biological Chemistry
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    • v.30 no.1
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    • pp.77-87
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    • 1987
  • The mutant with high productivity, X. malvacearum SNUF 560-6, was acquired from the X. malvacearum SNUF 560 with low productivity by UV-light irradiation. It was preserved is lyophilized stock culture and it was transferred to PDA slant to maintain viability fortnightly. Fermentations were started by retransfering to MY agar slant from PDA stok culture. The experiments for optimal xanthan gum production were studied in a chemically defined medium. Of the carbon and nitrogen sources tested, 0.4% sucrose medium and 10mM glutamic acid medium yielded the highest xanthan gun production respectively. The addition of 10g/l succinic acid stimulated xanthan gum production. Also 65mM $PO_4\;^{-3}\;(12.6g/l\;KH_2PO_4)$ was effective on xanthan gum production. Finally, medium 1 and medium 2 which have high xanthan gum production potencies were achieved in this stud. The components of medium 1 and medium 2 were as follows: Medium 1 : sucrose 40g/l glutamate 10mM $PO_4\;^{-3}\;54mM\;(KH_2PO_4\;12.65g/l)$ Citrate 2g/l $MgSO_4{\cdot}7H_2O\;0.2g/l$ $H_3BO_3\;0.005/l$ ZnO 0.006/l $FeCl_2{\cdot}6H_2O\;0.0024g/l$ $CaCO_3\;0.02g/l$ Medium 2 : $Sucrose\;40g/l\;(NH_4)_2SO_4\;2g/l$ $PO_4\;^{-3}\;65mM\;(KH_2PO_4\;12.65g/l)$ Succinate 10g/l $MgSO_4{\cdot}7H_2O\;0.02g/l$ $H_3BO_3\;0.06g/l$ ZnO 0.006g/l $FeCl_2{\cdot}6H_2O\;0.0024g/l$ $CaCO_3\;0.02g/l$.

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Isolation of Synthetic Detergent Decomposing Microorganisms in Wastewater and Synthetic Detergent Decomposition Characterization of the Microorganisms (폐하수중 합성세제분해균의 분리(分離) 및 합성세제 (ABS) 분해특성(分解特性))

  • Lee, Hong-Jae;Heo, Jong-Soo;Cho, Ju-Sik;Han, Mun-Gyu;Choi, Jeong-Ho;Lee, Chun-Hee
    • Korean Journal of Environmental Agriculture
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    • v.12 no.2
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    • pp.144-152
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    • 1993
  • A bacterium which degrades efficiently synthetic detergents was isolated from the polluted waters, activated sludge of wastewater treatment plants or polluted soil. This bacterium showed considerably higher growth rate in the agar plate containing $2,000{\mu}g/ml$ of synthetic detergents than any other isolated strains, was identified as a Pseudomonas fluorescens or strains similar to it. The strain was named as a Pseudomonas fluorescens S1. Optimum pH and temperature for the growth of the Pseudomonas fluorescens S1 were pH 7.0 and $30^{\circ}C$, respectively. The strain was resistant to streptomycin and gentamycin, but sensitive to kanamycin. The strain was greatly resistant to zinc chloride, lead nitrate and copper sulfate, but unable to grow in the presence of relatively low concentrations of mercury chloride and silver nitrate. This strain utilized benzene, catechol, cyclohexane and xylene as a sole carbon source. The strain was well grown in the medium containing ABS 10,000${\mu}g$/ml. Degradation of ABS was 55% and 60% at 20${\mu}g$/ml and 100${\mu}g$/ml of ABS, respectively. Benzene ring was degraded 45% in 100${\mu}g$/ml of ABS. During the incubation of the strain in the medium containing ABS 100${\mu}g$/ml and COD 10,000${\mu}g$/ml for 4 days, degradation of ABS and COD were reduced to 40${\mu}g$/ml and 3,200${\mu}g$/ml, respectively. Total amino acid content of the Pseudomonas fluorescens S1 grown with 1,000${\mu}g$/ml of ABS was 115mg/g cell, whereas its content was decreased in the bacterium grown without synthetic detergent by 9.4%.

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Isolation and Characterization of Exopolysaccharide Producing Lactic Acid Bacteria from Korean Soy Sauce and Soybean Paste (전통 장류로부터 Exopolysaccharide 생성 유산균의 분리 및 특성)

  • Yun, Hye Ju;Lee, You Jung;Yeo, Soo-Hwan;Park, Hye Young;Park, Heui-Dong;Baek, Seong Yeol
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.190-197
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    • 2013
  • Three slime-forming lactic acid bacteria were isolated from traditional Korean fermented soy sauce and soybean paste and shown to produce exopolysaccharides (EPS) in sucrose media. By isolating the strains, examining their morphological characteristics and determining their 16S rDNA sequences, N58-5 and K6-7 were identified as Leuconostoc mesenteroides and N45- 10 as Leuconostoc citreum. The acid and bile tolerances of these three strains were investigated. Amongst the three lactic acid bacteria, Leuc. citreum N45-10 exhibited the highest viability ($10^5-10^6$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 0.3) for 2 h, in artificial gastric juice for 2 h and in 0.3%, 0.5% oxgall for 24h. Leuc. mesenteroides K6-7, N58-5 and Leuc. citreum N45- 10 were grown in sucrose liquid medium and 8.16 g/L, 3.65 g/L, 16.17 g/L of EPS was collected, respectively. The hydrolyzed EPS was analyzed by HPLC in order to determine the sugar composition of EPS. Leuc. mesenteroides K6-7 and N58-5 showed two peaks indicating glucose and fructose, thus they were determined to be hetero-type polysaccharides. Leuc. citreum N45-10 showed only the glucose polymer, indicating it to be a homo-type polysaccharide. In addition, all three lactic acid bacterial hemolysis did not demonstrate a clear zone in blood agar in the area surrounding a lactic acid bacteria colony.