• Title/Summary/Keyword: Biolog^TM system

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Rapid Identification of Vibrio vulnificus in Seawater by Real-Time Quantitative TaqMan PCR

  • Wang, Hye-Young;Lee, Geon-Hyoung
    • Journal of Microbiology
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    • v.41 no.4
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    • pp.320-326
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    • 2003
  • In order to identify Vibrio vulnificus in the Yellow Sea near Gunsan, Korea during the early and late summers, the efficiency of the real-time quantitative TaqMan PCR was compared to the efficiency of the conventional PCR and Biolog identification system^TM. Primers and a probe were designed from the hemolysin/cytolysin gene sequence of V. vulnificus strains. The number of positive detections by real-time quantitative TaqMan PCR, conventional PCR, and the Biolog identification system from seawater were 53 (36.8%), 36 (25%), and 10 strains (6.9%), respectively, among 144 samples collected from Yellow Sea near Gunsan, Korea. Thus, the detection method of the real-time quantitative TaqMan PCR assay was more effective in terms of accuracy than that of the conventional PCR and Biolog system. Therefore, our results showed that the real-time TaqMan probe and the primer set developed in this study can be applied successfully as a rapid screening tool for the detection of V. vulnificus.

A Data Base for Identification of Pseudomonas syringae pv. actinidiae, the Pathogen of Kiwifruit Bacterial Canker, Using Biolog Program (Biolog Program을 이용한 참다래 궤양병균 동정용 Data Base)

  • 고영진
    • Korean Journal Plant Pathology
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    • v.13 no.2
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    • pp.125-128
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    • 1997
  • Reactions of Pseudomonas syringae pv. actinidiae to 95 carbon sources in a 96-well microplate (BiOLOG GN MicroPlateTM) were investigated. The bacterium used 9 carbon sources such as D-mannitol, sucrose, etc., but did not use 62 carbon sources such as $\alpha$-cyclodextrin, dextrin, etc. Based on the reactions, a user data base for identification of P. syringae pv. actinidiae was constructed in Biolog program (BiOLOG MicroLogTM 2 system). P. syringae pv. actinidiae isolates collected from kiwifruits could be identified automatically with high similarity using the user data base, which could diagnose rapidly and easily whether the tree was infected with bacterial canker or not.

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Hydroxyapatite Solubilization and Organic Acid Production by Enterobacter agglomerans (인산염 가용화균 Enterobacter agglomerans에 의한 Hydroxyapatite 가동화와 유기산 생성)

  • Kim, Kil-Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.30 no.2
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    • pp.189-195
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    • 1997
  • A phosphate solubilizing bacterium (PSB) possessing a high ability to solubilize hydroxyapatite (HA) was isolated from the rhizosphere of wheat. The PSB markedly developed clear zones after inoculating for 36 hours at $30^{\circ}C$. This bacterium was identified as Enterobacter agglomerans through API 20E system and Biolog$^{TM}$ analysis. The values of similarity and distance coefficient from authentication trial of the strain were 0.656 and 4.79 respectively. High performance liquid chromatography (HPLC) of the products of this strain indicated that this strain excretes maily oxalic acid with som other organic acids. During the incubation period of E. agglomerans, the pH values showed an inverse correlation ($r^2=0.933^{**}$) with solubilization of inorganic phosphate. Acid phosphatase activity of the strain was 10-15 times greater than alkaline phosphatase activity. Alkaline phosphatase activity had almost constant near zero activity across time. The population of E. agglomerans greatly increased during the first day of inoculation ; however, it drastically decreased thereafter.

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Isolation and Identification of the Antagonistic Microorganisms Against Streptococous spp. Causing Dental Caries in Korean Soy Sauce (한국재래간장으로부터 구강질환 방제균의 선발 및 동정)

  • 엄수정;이여진;김진락;이은탁;김상달
    • Journal of Life Science
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    • v.13 no.4
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    • pp.535-540
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    • 2003
  • The antagonistic microorganisms against Streptococcus sanguis, S. salivarius and S. mutans causing the dental caries of oral diseases were isolated from Korean traditional soy sauce. Twenty five strains were isolated by pairing culture, paper disc culture and dual culture methods. The isolate NG 06 strain was observed with various cultural and physiological test, and $Biolog^{(R)}$ Bacterial Identification System. The strain was identified as Bacillus racemilacticus. The isolate NG 16 strain was confirmed to Gram-positive, rods, endospore production, utilization of melibiose, casein hydrolysis and starch hydrolysis. Also the second strain NG 16 was identified as $\beta$. amyloliquefaciens.

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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Bacteriological Characteristics of Unidentified Vibrio sp., Hemolysin Producer Isolates front Brackish Water -1. Bacteriological Characteristics of Vibrio sp., D9 (V. kumkang) Similar to V. mimicus (기수에서 분리된 용혈독소를 생산하는 미분류 Vibrio sp.의 세균학적 특징 -1. V. mimicus와 유사한 Vibrio sp. D9의 세균학적 특성)

  • KIM Young-Man;OH Hee-Kyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.6
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    • pp.585-590
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    • 2000
  • A hemolysin producing strain was isolated from Kum rivet estuary located in west part of Korea. In the process of identification the isolated strain was similar to V. mimicus but did not show characteristics of known Vibrio species; therefore, the strain was designated as Vibrio sp. D9 ( V. kumkang) tentatively and further identification study was carried out by comparing its bacteriological characteristics, Morphologically Vibrio sp. D9 was a typical straight roe with a polar flagellium. Among known Vibrio species no identical strains were found when using automatic bacteria identification system ($MicioLog^(TM)$system, release 4.0, Biolog Inc., USA) which evaluated the ability of metabolizing 95 kinds of carbon and nitrogen sources. Vibrio sp. D9 showed 18 and 13 different responses as compared to V. mimicus and V. cholerae, respectively. Clear hemolysis zones were observed with the strain against human and sheep blood agar plate, Hemolytic toxicity was confirmed by strong vascular permeability and fatal toxicity against mouse was also observed. Thus the strain was a pathogenic vibrio. Growth conditions for Vibrio sp. D9 were salinity of $0{\~}5.0{\%}$, pH of $6.4{\~}9.8$, temperature of $15{\~}41^{\circ}C$, respectively.

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Chemical Properties of Pine Sprout Tea and Identification of the Related Yeasts (송순차의 화학적 특성 및 품질관련 효모의 동정)

  • Kang, Jeong-Hwa;Yoo, Maeng-Ja;Chung, Hee-Jong
    • Journal of the Korean Society of Food Culture
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    • v.15 no.4
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    • pp.233-239
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    • 2000
  • In order to optimize the quality of pine sprout tea, its chemical properties were analyzed and the yeasts associated with the quality of pine sprout tea during the storage were isolated and identified. In proximate composition moisture content was 20.13%, but other components except sugar were relatively low. Sugars such as glucose(30.15%), fructose(19.57%), and sucrose(9.27%) were major sugars which contained up to 76.73%. Total vitamin C and soluble tannin contents were 11.31 mg% and 68.31 mg%, respectively. Thirteen kinds of free amino acids were detected, but they were contained only in trace. In fatty acid composition 64.69% of fatty aids composed mainly of saturated fatty acids and major fatty acids were oleic acid, palmitic acid, and tricosaenoic acid. Among 8 mineral elements detected, calcium content was highest with 79.00 mg% and followed by potassium(45.16 mg%) and magnesium(8.93 mg%). The sweetness of pine sprout tea was gradually decreased from $70^{\circ}\;Brix\;to\;63^{\circ}\;Brix$ and 3.2% of ethanol at the initial concentration was increased to 6.0% during the storage of 40 days. The yeasts associated with the quality and alcohol formation of pine sprout tea during the storage were identified by Biolog MicrostationlTM system, as Zygosaccharomyces rouxii, Kluyveromyces lodderae, Kluyveromyces wickerhamii, and Pichia fluxuum.

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Isolation, Identification and Cultural Condition of the Antagonistic Microorganism Against Salmonella gallinarum Causing Fowl Typhoid (가금티브스균 Salmonella gallinarum의 생육을 저해하는 길항미생물의 선발 및 동정)

  • 김진락;김상달
    • Journal of Life Science
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    • v.13 no.6
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    • pp.843-848
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    • 2003
  • Diarrllea and death of chicken have been brought about by fowl typhoid caused by Salmonella gallinarum, which causes a great loss of chicken farms. For the development of the probiotic which can control a fowl typhoid of S. gallinarum without any adverse effect of commercial existing antibiotics, we isolated antagonistic intestinal bacteria against S. gallinarum from a bowel of the chicken which was pastured in a chicken farm of Gumi, Kyoungbuk. An Y3 strain which had a strong antagonistic ability to S. gallinarum was selected as a candidate of chicken probiotic microorganism among isolated strains. It was identified as a Bacillus amyloliuefaciens by 98% similarity by the result of cultural, physiological, biochemical test and Biolog system$(Microlog^{TM} 4.0)$, and named as Bacillus amyloliquefaciens Y3. The strain showed the strongest antagonistic activity and a good growth at pH 5-9, $37^{\circ}C.$

Bacteriological Characteristics of Unidentified Vibrio sp., Hemolysin Producer Isolated from Brackish Water -2. Bacteriological Characteristics of Vibrio sp. E10 Similar to Vibrio mimicus- (기수에서 분리된 용혈독소를 생산하는 미분류 Vibrio sp.의 세균학적 특징 -2. Vibrio mimicus와 유사한 Vibrio sp. E10의 세균학적 특성-)

  • Kim Young Man;Yu Hong Sik;Oh Hee Kyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.545-550
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    • 2002
  • A hemolysin producing bacterial strain which belong to Vibrio species was isolated from the Kum River estuary. In the process of identification, the strain did not show characteristics of known Vibrio species; thus, the strain was designated as Vibrio sp, E10 (V. kunsan) tentatively and further identification study was carried out by comparing its bacteriological characteristics. Morphologically Vibrio sp, E10 was comma shaped rod with a polar flagellium. Clear hemolysis zones were observed with the strain against human and sheep blood agar. Hemollytic toxicity was confirmed by strong vascular Permeability and fatal toxicity against mouse was also observed. Therefore the strain was a pathogenic vibrio. Growth conditions for Vibrio sp. E10 were ranged salinity of 0$\~$$4.5\%$, pH of 6.2$\~$9.2, temperature of 14$\~$42$^{\circ}C$, respectively, 16S rDNA partial sequence of Vibrio sp, E10 showed $99\%$ homology with dozens of V. cholerae species including V, cholerae El Tor N16961 and V, snmisnfus ATCC 33653T. This strain belonged to Proteobacteria; gamma subdivision; Vibrionacea: Vibrio. But, among knorn Vibrio species no identical styains were found when using automatic bacteria identification system ($MicroLog^{TM}$system, release 4.0, Biolog Inc., USA) which evaluated the ability of metabolizing 95 kinds of carbon and nitrogen sources. Vibrio sp, E10 showed 18 and 11 different responses as compared to V. mimicus and V, cholerae, respectively.

A Multi-microbial Biofungicide for the Biological Control against Several Important Plant Pathogenic Fungi (진균성 식물병해 방제를 위한 항생물질 생산 길항미생물의 복합제제화)

  • Jung, Hee-Kyoung;Ryoo, Jae-Cheon;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.48 no.1
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    • pp.40-47
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    • 2005
  • In order to develop a multi-microbial biofungicide against several important plant pathogenic fungi, strains were isolated from the phtophthora blight suppressive red-pepper field soil of Gyeongsangbuk-do, Korea. Strains AY1, AY6, AB1, BB2 and F4, which had strong antagonistic ability against Phytophthota capsici and Fusarium oxysporum, were selected for their involvement with strains of biocontrol fungicide. There were no antagonism among the selected strains and were compatible for making the biofungicide. Their antagonistic mechanisms, except for strain BB2, were an antibiosis by the production of antibiotic, while BB2 produced not only an antibiotic but also cellulase as an antagonistic mechanism against blight causing P. capsici. They were identified as Halobacterium sp. AB1, Xenorhadus sp. AY1, Bacillus sp. AY6, Bacillus sp. BB2, Zymomonas sp. F4 by various cultural, biochemical test and $Biolog^{TM}$ System 4.0. The highest levels of antifungal antibiotic could be produced after 48 hrs of incubation under the optimal medium which were 0.1% galactose, 0.1% $NaNO_2$, 5 mM $Na_2{\cdot}HPO_4$ (pH 5.5). The cultured multi-microbial biofungicide showed strong biocontrol activity against bacterial wilt disease and fusarium wilt disease in cucumber and tomato fields.