• Title/Summary/Keyword: Selective medium

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Identification of Lactobacillus ruminus SPM0211 Isolated from Healthy Koreans and Its Antimicrobial Activity against Some Pathogens

  • Yun Ji-Hee;Yim Dong-sool;Kang Jin-Yang;Kang Byung-Yong;Shin Eun-ah;Chung Myung-Jun;Kim Soo-Dong;Baek Dae-Heoun;Kim Kyungjae;Ha Nam-Joo
    • Archives of Pharmacal Research
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    • v.28 no.6
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    • pp.660-666
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    • 2005
  • The intestinal microbiota are important to the host with regard to resistance they impart against bacterial infections and their involvement in mediating metabolic functions. Lactic acid producing bacteria such as Lactobacillus play an important physiological role in these matters. The aim of the present study was to isolate Lactobacillus sp. that inhibits enteric pathogens. Initially, 17 isolates from healthy Koreans were collected on Lactobacillus selective medium. Resistance of the isolates to antibiotics including rifampicin, streptomycin, clindamycin and vancomycin was measured. One of the isolate was identified as Lactobacillus ruminus on the basis of bacterial cell morphology, cultural characteristic and biochemical characteristics, 16S rRNA sequence analysis and PCR-RAPD. Antimicrobial activity of the bacterium against Vancomycin Intermediate Resistant Staphylococcus aureus (VISA) and Vancomycin-Resistant Enterococci (VRE) was measured. About $10^4$ cells of VISA or VRE were mixed with 1, 5, and 9 mL of L. ruminus SPM 0211 and the final volume was adjusted to 10 mL with brain heart infusion (BHI) broth. The cell suspension was incubated for 3, 6, 9, and 24 h, serially diluted and then plated on BHI agar plates. As numbers of L. ruminus SPM 0211 were increased, viable cell count of VISA and VRE decreased. The strongest antimicrobial activity of SPM 0211 was observed after 9 h incubation in any mixture, almost completely inhibiting the growth of these two bacteria. The results suggest that the freshly isolated L. ruminus SPM 0211 may be used as a pro-biotic microbe that prevents the colonization of enteric pathogens and can thereby promote good gastrointestinal health.

Chestnut Ink Disease Caused by Phytophthora katsurae (Phytophthora katsurae 에 의한 밤나무 잉크병)

  • Oh, E.;Lee, J.-K.;Lee, S.-H.;Kim, K.-H.
    • Journal of Forest and Environmental Science
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    • v.23 no.1
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    • pp.65-71
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    • 2007
  • In early 2000's, about six Phytophthora species have been newly described leading mortality on coniferous and broad-leaved trees in forests. Also, some species of Phytophthora are responsible for ink disease in chestnut plantation near or within forests. Similar symptoms of ink disease were appeared in some areas of Kyungnam and Jeonnam providences in 2005, and the pathogen was isolated using Phytophthora- selective medium in 2006. Morphological and genetic analysis were performed to identify the isolate. Also, the pathogenicity was conducted to complete $K\ddot{o}ch^{\prime}s$ postulate and compare susceptibility among chestnut cultivars. The molecular analysis between P. katsurae and P. hevae were performed with the isolates obtained from different countries including Korea or the sequences downloaded from Phytophthora webpage. The result showed that the isolated pathogen from chestnut was P. katsurae. There is no report of P. katsurae in Korea until now. P. katsurae was re-isolated from inoculated chestnut cultivars. Also, there was a slight difference in susceptibility among chestnut cultivars. The rDNA sequence of our isolate showed 100% similarity with sequence of the isolate cultured from Japan and New Zealand.

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Voltammetric Determination of Copper(II) Using Glassy Carbon Electrodes Modified with Nafion-DTPA-Glycerol

  • Park, Chan-Ju;Park, Eun-Heui;Chung, Keun-Ho
    • Proceedings of the Korean Environmental Health Society Conference
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    • 2003.06a
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    • pp.177-180
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    • 2003
  • A glassy carbon electrode(GCE) modified with nafion-DTPA-glycerol was used for the highly selective and sensitive determination of a trace amount of Cu$\^$2+/. Various experimental parameters, which influenced the response of nafion-DTPA-glycerol modified electrode to Cu$\^$2+/, were optimized. The copper(II) was accumulated on the electrode surface by the formation of the complex in an open circuit, and the resulting surface was characterized by medium exchange, electrochemical reduction, and differential pulse voltammetry, A linear range was obtained in the concentration range 1.0${\times}$10$\^$-8/M∼1.0${\times}$10$\^$-6/M Cu(II) with 7 min preconcentration. Further, when an approximate amount of lead(II) is added to the test solution, nafion-DTPA-glycerol modified glassy carbon electrode has a dynamic range of 2 orders magnitude(1.0${\times}$10$\^$-9/M∼1.0${\times}$10$\^$-7/M). The detection limit(3 $\sigma$) was as low as 5.0${\times}$10$\^$-6/M(0.032ppb). The interferences from other metal ions could be reduced by adding KCN into the sample solutions. This method was applied to the determination of coppe,(II) in certified reference material(3.23${\times}$10$\^$-7/M, 21ppb), sea water(9.50${\times}$10/sup-7/M, 60ppb). The result agrees satisfactorily with the value measured by Korea Research Institute of Standard and Science.

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INTERRELATIONSHIP BETWEEN VIRULENT CLONAL TYPES, SEROTYPES AND LEUKOTOXICITY OF KOREAN STRAINS OF A. ACTINOMYCETEMCOMITANS (한국인 Actinobacillus actinomycetemcomitans 균주의 특이 독성 clone형과 혈청형 및 백혈구독성과의 관계)

  • Ku, Young
    • Journal of Periodontal and Implant Science
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    • v.25 no.3
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    • pp.487-496
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    • 1995
  • Previous studies have demonstrated that not all A. actinomycetemcomitans produced significant level of leukotoxic factor and its leukotoxicity have associated with serotype and genetic variation. Our aim was to investigate on the interrelationship between serotype and leukotoxicity of an A. actinomycetemcomitans consisting of 13 clinically well characterized. Korean isolates and to evaluate if particular virulent clonal types of A. actinomycetemcomitans are associated with periodontal disease. For this study, 13 strains of A. actinomycetemcomitans from 6 patients with periodontal disease were isolated and identified by using a selective medium(tryptic soy agar supplemented with 10% serum, $75{\mu}g$ of bacitracin and $5{\mu}g$ of vancomycin per ml) in 10% C02 incubator for 3days with routine Gram staining, colony morphology and biochemical test..For serotyping, antisera were prepared from reference strains of 5 serotypes. (ATCC 29523,Y4, SUNY aB 67, IDH 781, IDH 1705) and then ammonium sulfate precipitation, immunoabsorption and indirect immunofluoroscent procedures were done. For analysis of leukotoxicity, sonic extract of A. actinomycetemcomitans exposed to PMN, and trypan blue was stained for counting the cell viability. Finally Southern blot analyses of genomic DNA digested with the restriction enzyme Tag I was done and the Southern blots were hybridized with the 530bp fragment, termed delta 530, originating from the ltx promoter of strain 652 and deleted from strain JP2. Also ltxA-3.1 and SC2 probe from strain JP2 were hybridized with genomic DNA fragments. Results reveal that strains isolated showed approximately equal proportions of 3 serotypes(b, d, e) and serotype b was not detected. 2 patients harbored 2 different serotypes in the same disease site. The prevalence of leukotoxic strain was 23% and there was no relationship between serotype, leukotoxicity and clinical observations. Especially virulent clonal types of Actinobacillus actinomycetemcomitan (JP2 strain) could not found. Further studies are necessary on the genetic polymorphism of leukotoxin and its relations to clinical status.

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GUS gene expression and plant regeneration via co-culturing with Agrobacterium in grapevine (Vitis vinifera) (Agrobacterium 공동배양을 이용한 포도 재분화율 향상과 GUS 유전자의 발현)

  • Kim, Se-Hee;Kim, Jeong-Hee;Kim, Ki-Ok;Do, Gyeong-Ran;Shin, Il-Sheob;Cho, Kang-Hee;Hwang, Hae-Seong
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.308-314
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    • 2011
  • Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. In this study, methods for Agrobacterium tumefaciens-mediated transformation and plant regeneration of grapevine (Vitis vinifera) were evaluated. Tamnara, Heukgoosul, Heukbosek, Rizamat were co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, GUS gene as reporter gene and resistance to kanamycin as selective agent. Seven percent of the maximum regeneration frequency was obtained from co-cultivated with explants from Rizamat with LBA4404 strain on selection medium with kanamycin. The addition of acetosyringone, 200 ${\mu}m$ in virulence induction step was a key factor for successful GUS reporter gene expression in grapevine transformation. Transgenic plants showed resistance to kanamycin and the GUS positive response in leaf ($T_0$) stem ($T_0$) and petiole ($T_0$).

Effects of Temperature and Moisture on the Survival of Colletotrichum acutatum, the Causal Agent of Pepper Anthracnose in Soil and Pepper Fruit Debris

  • Kang, Beum-Kwan;Kim, Joo-Hyeong;Lee, Kyeong-Hee;Lim, Sang-Cheol;Ji, Jae-Jun;Lee, Jong-Won;Kim, Heung-Tae
    • The Plant Pathology Journal
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    • v.25 no.2
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    • pp.128-135
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    • 2009
  • The survival of Colletotrichum acutatum was investigated in soil, infected fruits, and infected fruit debris incorporated into soil at several temperatures with different soil moisture levels. Samples were examined at 2-week intervals for 18 weeks to determine the survival of the pathogen based on the number of colony forming unit (CFU) of C. acutatum recovered on a semi-selective medium. C. acutatum conidia survived in both sterile and non-sterile soil at 4 and $10^{\circ}C$ for 18 weeks. If infected pepper fruits were completely dried, C. acutatum survived for 18 weeks at temperature from 4 to $20^{\circ}C$. Soil temperature and moisture affected the survival of C. acutatum in infected fruit debris incorporated into soil after air-drying. The effect of soil moisture on survival was weaker at low temperatures than at high temperatures. For up to 16 weeks, conidia were recovered from fruit debris in soil that had been kept at 4 to $20^{\circ}C$ and below 6% soil moisture. Conidia were recovered from fields until approximately 6 months after pepper fruits were harvested. Using PCR with species-specific primers and a pathogenicity test, we identified conidia recovered from soil and infected fruit from both the laboratory and field as C. acutatum and as the primary inoculum causing pepper anthracnose.

Plant quarantine isolated cultivation system in Korea and results of recorded in 2005-2012 (우리나라 식물검역 격리재배 시스템과 2005-2012년 실적보고)

  • Lee, Siwon;Park, Jungan;Lee, O-Mi;Shin, Yong-Gil
    • Korean Journal of Agricultural Science
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    • v.40 no.4
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    • pp.281-287
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    • 2013
  • In Korea, isolated cultivation has been implemented for 102 genera, including about 250 species, each of which has underwent microscopic inspection, cultivation of bacteria in selective medium, analysis of physiology and biochemistry, enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). The number of isolated microorganisms was 8,307 in the period of 2005-2012, and bulbs and tubers had the greatest diversity of microorganisms, of 5,165 (62.2%), followed by 2,119 (25.0%) sapling, 796 (9.6%) seed, 150 (1.8%) cutting slip, 70 (0.8%) branch graft and 7 (0.1%). The number of cases which were disqualified were 413 (4.97%), after the detection of 47 disease causing species of microorganism. Viruses predominated, with 27 species, followed by 16 fungi, a viroid, a Chromalveolata and 2 further species. Top on the list of detection was Arabis mosaic virus (77 cases), followed by Tobacco rattle virus (70 cases), Lily symptomless virus (46 cases) and Penicillium expansum (46 cases).

Growth Modelling of Listeria monocytogenes in Korean Pork Bulgogi Stored at Isothermal Conditions

  • Lee, Na-Kyoung;Ahn, Sin Hye;Lee, Joo-Yeon;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.108-113
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    • 2015
  • The purpose of this study was to develop predictive models for the growth of Listeria monocytogenes in pork Bulgogi at various storage temperatures. A two-strain mixture of L. monocytogenes (ATCC 15313 and isolated from pork Bulgogi) was inoculated on pork Bulgogi at 3 Log CFU/g. L. monocytogenes strains were enumerated using general plating method on Listeria selective medium. The inoculated samples were stored at 5, 15, and $25^{\circ}C$ for primary models. Primary models were developed using the Baranyi model equations, and the maximum specific growth rate was shown to be dependent on storage temperature. A secondary model of growth rate as a function of storage temperature was also developed. As the storage temperature increased, the lag time (LT) values decreased dramatically and the specific growth rate of L. monocytogenes increased. The mathematically predicted growth parameters were evaluated based on the modified bias factor ($B_f$), accuracy factor ($A_f$), root mean square error (RMSE), coefficient of determination ($R^2$), and relative errors (RE). These values indicated that the developed models were reliably able to predict the growth of L. monocytogenes in pork Bulgogi. Hence, the predictive models may be used to assess microbiological hygiene in the meat supply chain as a function of storage temperature.

Identification of Agrobacterium tumefaciens from Soil and Transformation of Maize (토양으로부터의 Agrobacterium tumefaciens의 분리, 동정 및 옥수수의 형질전환에 이용)

  • 노광수;강봉중
    • KSBB Journal
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    • v.7 no.3
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    • pp.191-200
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    • 1992
  • Several strains of Agrobacterium tumefaciens were isolated from soil in the Taegu area and characterized to develop some useful vector systems for higher plant genetic engineering. The selected colonies had a unique form, and strains from the colonies were capable of tumor formation on the sunflower leaf surface. They had a large plasmid. The restriction analysis showed that they were another kinds of Ti plasmic compared with C58 and Ach5. The isolated strains were identified as the nopaline type and also as biovar 1 A. tumefaciens, according to their tumor morphology, blophyslcal and biochemical characteristics. One of the isolated strains, AK204 was transformed with binary vector (pGA642), having selectable marker (Kmr, Tcr). Furthermore, maize tissue cells were transformed by cocultivation with AK204/pGA642, and the transformants were selected on the selective medium and identified using PAGE patterns of their soluble proteins.

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Analysis of cellular fatty acid methyl esters (FAMEs) for the identification of leuconostoc strains isolated from kimchi

  • Lee, Jung-Sook;Chun, Chang-Ouk;Kim, Hong-Joong;Joo, Yun-Jung;Lee, Hun-Joo;Park, Chan-Sum;Park, Yong-Ha;Mheen, Tae-Ick
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.225-228
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    • 1996
  • The cellular fatty acid methyl esters (FAMEs) analysis data obtained for clusters defined at a Euclidian distance of 17.5, in the classification of lactic acid bacteria isolated from kimchi, described by Lee et al. (4), was used for the identification of 79 Leuconostoc isolates. The test strains were isolated using a selective isolation medium specific for the genus Leuconostoc. These strains were then characterized according to their fatty acid profiles. The results show that all seventy nine test strains were identified to the known Leuconostoc clusters B, C, and D. Cluster B had the highest relative amount of the saturated fatty acid 16 : 0. The saturated fatty acid 16 : 0 and summed feature 9 were found as a major components in cluster C, which had a higher level of summed feature 9 than cluster B. Cluster D is characterized by the highest relative amount of the unsaturated fatty acid 18 : 1 w9c. It is suggested that FAMEs analysis can be successfully applied in the identification of lactic acid bacteria isolated from kimchi.

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