• Title/Summary/Keyword: E-coli

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Seroprevalence of Brucellosis and Isolation of Yersinia enterocolitica O:9 in Pigs (돼지에서 브루셀라병 항체조사 및 Yersinia enterocolitica O:9의 분리)

  • Jung, Byeong-Yeal;Byun, Jae-Won;Kim, Ha-Young;Shin, Dong-Ho;Park, Choi-Kyu;Jung, Suk-Chan
    • Journal of Life Science
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    • v.20 no.5
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    • pp.697-702
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    • 2010
  • Ten farrow-finish farms participated in this seromonitoring that was conducted to investigate the porcine brucellosis situation in Korea. In total, eight (80.0%) of the 10 farms and 139 (24.0%) of 578 pigs tested showed a positive response in the Rose Bengal test (RBT). Seroprevalence levels were determined using RBT according to age; 35 (14.6%) of 239 piglets, 36 (31.3%) of 115 growing pigs, and 68 (30.4%) of 224 finishing pigs and sows were positive, respectively. All positive samples in RBT were tested with the tube agglutination test (TAT) and competitive ELISA (C-ELISA), simultaneously. Although 48 samples came up positive in the TAT, all samples tested with C-ELISA were negative. Among 26 rectal swab samples from the TAT positive-pigs, Yersinia enterocolitica O:9 was isolated from seven samples (26.9%). Therefore, we speculated that the positive reaction of RBT and TAT in this study might be induced by the serologically cross-reacting bacteria with Brucella abortus.

Functional Role of Peptide Segment Containing 1-25 Amino Acids in N-terminal End Region of ErmSF (ErmSF에서 특이적으로 발견되는 N-terminal end region에 존재하는 1-25번째 아미노산을 함유하는 peptide segment의 효소 활성에서의 역할)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.165-171
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    • 2006
  • ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA to confer the resistance to MLS (macrolide-lincosamide-streptogramin B) antibiotics on microorganism ranging from antibiotic producers to pathogens. To define the functional role of peptide segment encompassing amino acid residues 1 to 25 in NTER (N-terminal end region) of ErmSF, one of the ERM proteins, DNA fragment encoding mutant protein deprived of that peptide was cloned and overexpressed in E. coli to obtain a purified soluble form protein to the apparent homogeneity in the yield of 12.65 mg per liter of culture. The in vitro activity of mutant protein was found to be 85% compared to wild type ErmSF, suggesting that this peptide interact with substrate to affect the enzyme activity. This diminished activity of mutant protein caused the delayed expression of antibiotic resistance in vivo, that at fIrst cells expressing mutant protein showed the retarded growth due to the antibiotic action but with time cells inhibited by antibiotic gradually recovered the viability to exert the resistance to the same extent as those with wild type protein.

Quality Changes of Egg Products during Storage (계란가공품의 보관중 품질 변화)

  • Kim, Jae-Wook;Kim, Hyong-Chan;Hur, Jong-Wha
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1480-1483
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    • 1998
  • To know the probable storage conditions for commercially prepared ‘baked egg’ and ‘rolled egg’ products, microbes and quality changes for samples immediately after the production and stored under the several conditions were tested. Sealed samples were stored at 5, 10, 15, $20^{\circ}C$, respectively and tested at 1 or 2-days intervals, opened samples were stored at 15, 20, 30, $37^{\circ}C$, respectively and tested at 6-hr intervals. Coliform bacteria, E. coli, Salmonella, Staphylococci were not detected, and viable cell counts level was under $10^2\;CFU/g$ for samples immediately after the production. The higher the storage temperature and baked eggs rether than rolled eggs showed more rapid microbial growth for both samples. For sealed condition, 14-days at $5^{\circ}C$ for both samples, 7-days for rolled eggs and 5-days for baked eggs at $10^{\circ}C$ were kept their initial microbial level and favorable flavor. For opened condition, it showed remarkable microbial increase after $18{\sim}24\;hr$ for $15{\sim}20^{\circ}C$ and 12 hr for $30{\sim}37^{\circ}C$. From these results, used egg products samples are able to be stored more than 1-week at below $5^{\circ}C$ and opened samples need to be stored at cold storage. At room temperature, it egg products.

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Antimicrobial Activity of Solvent Fraction from Cornus officianalis (산수유의 용매분획별 항균활성)

  • 김용두;김황곤;김경제
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.6
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    • pp.829-832
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    • 2003
  • To develop natural food preservatives, ethanol and water extracts were prepared from the cornus (Cornus of officianalis) and antimicrobial activities were examined against 10 microoganisms which were food borne pathogens and/or food poisoning microoganisms, food-related bacteria and yeasts. Ethanol extract exhibited antimicrobial activity for the microoganisms tested, except lactic acid bacteria and yeast. Especially, minimum inhibitory concentrations (MIC) of the ethanol extracts were determined as 0.25 mg/mL against bacteria and 2 mg/mL against target lactic bacteria and yeasts. Antimicrobial activity of the ethanol extracts were not destroyed by the heating at 121$^{\circ}C$ for 15 min and not affected by pH. The ethanol extract of cornus exhibiting high antimicrobial activities were fractionated in the other of diethylether and butanol fractions to test antimicrobial activity The antimicrobial activity adjust bacteria test was highest in the ethanol fraction.

A Study on the Ecological Restoration Strategies for the Disturbed Landscapes (경관훼손지의 생태적 복구방안에 관한 연구)

  • Kim, Nam-Choon
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.1 no.1
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    • pp.28-44
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    • 1998
  • This study was conducted to suggest the ecological restoration strategies for the disturbed landscapes by theoretical study. Especially, it is aimed to suggest three objectives for restoration by using native plants ; (1) prevention or reduction of wind and water erosion, (2) provision of food and cover for variety of animal species, (3) improvement of the visual or aesthetic quality of disturbed sites. The main results were summarized as follows. 1. Cooperation between restoration ecologists and restoration practitioners is needed to delineate a scientific approach to restoration ; (1) Being aware of published literature that describes similar work and/or establish general principles. (2) Preparedness to carry out proper experiments to test ideas. (3) Preparedness to monitor fundamental parameters in a restoration scheme. (4) Providing information about the behavior of species. (5) Publish results. 2. There are three models of succession in theories of plant succession. The tolerance and facilitation models were recommended to ecological restoration. The inhibition model applies in most secondary succession, but the actual species which are introduced first may inhibit the germination and growth of slower-growing species, or they may prevent the growth of other species whose propagules arrive later. 3. The objectives of erosion control, wildlife habitat provision, and visual quality improvement are not mutually exclusive. However, many revegetation practices in the past have emphasized one of these aspects at the expense of the others. 4. A native plant community can be the model of ecological restoration. By stylization/abstraction of native plant community, trying to learn the most essential characteristics of community types - environmental factors ; dominant, prevalent, and "visual essence" species composition - in order to use such information in restoration. 5. After developing mass/spaces plan, match plant communities to the mass/space plan. In utilizing community grouping, there needs aesthetic ability to understand design elements. 6. Several hydrophytes such as Pennisetum saccharifluous, Themeda triandra, Cirsium pendulum show relatively good germination rates. In case of mesophytes and xerophytes, Arundinella hina, Artemisia princeps, Oenothera odorata and legumes seem to have quick-germinating abilities at barren sites. Pinus thunbergii, Rhus chinensis, Evodia daniellii, Alnus firma and Albizzia julibrissin can be considered as "late succession" woody plants because they show low germinating rates and slow growing habitat. 7. The seeds used for restoration should be collected within a certain radius of where it will be planted. Consideration in genetic issues in the collection and use of germplasm can increase the odds for successful restoration efforts. 8. The useful model in the "drift" pattern occurs so abundantly in naturally evolving landscapes. As one species diminishes in density, a second or third species are increased. Thus, dynamic interactions between species are created. Careful using of "drift phenomenon" in planting was recommended. 9. Virtually no stand of vegetation today is immune from the introduction and/or spreading of exotic species. Therefore, the perpetuation of a restored sites requires conscious monitoring and management. Thus, management would be the most important process in ecological restoration. 10. In order to keep the sites "natural", alternative management strategies would be applied in management ; Atenative management strategies are decelerating successional process, accelerating successional processes or modifying species composition. As management tools, mechanical, chemical, biological or environmental factor manipulation, e.g., fire could be used.

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High-Level Expression of T4 Endonuclease V in Insect Cells as Biologically Active Form

  • Kang, Chang-Soo;Son, Seung-Yeol;Bang, In-Seok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1583-1590
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    • 2006
  • T4 endonuclease V (T4 endo V) [EC 3. 1. 25. 1], found in bacteriophage T4, is responsible for excision repair of damaged DNA. The enzyme possesses two activities: a cyclobutane pyrimidine dimer DNA glycosylase (CPD glycosylase) and an apyrimidic/apurinic endonuclease (AP lyase). T4 denV (414 bp cDNA) encoding T4 en do V (138 amino acid) was synthesized and expressed using either an expression vector, pTriEx-4, in E. coli or a baculovirus AcNPV vector, pBacPAK8, in insect cells. The recombinant His-Tag/T4 endo V (rHis-Tag/T4 endo V) protein expressed from bacteria was purified using one-step affinity chromatography with a HiTrap Chelating HP column and used to make rabbit anti-His-Tag/T4 endo V polyclonal antibody for detection of recombinant T4 endo V (rT4 endo V) expressed in insect cells. In the meantime, the recombinant baculovirus was obtained by cotransfection of BacPAK6 viral DNA and pBP/T4 endo V in Spodoptera frugiperda (Sf21) insect cells, and used to infect Sf21 cells to overexpress T4 endo V protein. The level of rT4 endo V protein expressed in Sf21 cells was optimized by varying the virus titers and time course of infection. The optimal expression condition was set as follows; infection of the cells at a MOI of 10 and harvest at 96 h post-infection. Under these conditions, we estimated the amount of rT4 endo V produced in the baculovirus expression vector system to be 125 mg/l. The rT4 endo V was purified to homogeneity by a rapid procedure, consisting of ion-exchange, affinity, and reversed phase chromatographies, based on FPLC. The rT4 endo V positively reacted to an antiserum made against rHis-Tag/T4 endo V and showed a residual nicking activity against CPD-containing DNA caused by UV. This is the first report to have T4 endo V expressed in an insect system to exclude the toxic effect of a bacterial expression system, retaining enzymatic activity.

Regulation of sfs1 gene expression by the cAMP-cAMP receptor protein (sfs1 유전자의 cAMP-cAMP receptor protein에 의한 발현 조절)

  • Yoo, Ju-Soon;Lee, Seung-Jin;Lee, Hee-Young;Chung, Soo-Yeol;Choi, Yong-Lark
    • Applied Biological Chemistry
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    • v.39 no.3
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    • pp.195-199
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    • 1996
  • We have cloned several E. coli sfs genes which stimulate mal gene expression with $crp^{{\ast}1}$). One the genes (pPVC2) was sequenced and potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. In order to investigate the regulation of the sfs1 gene by the cAMP-CRP complex, we have constructed the sfs-lacZ fusion gene in this research. The overall transcriptional stimulations of sfs1 gene in the presence cAMP were confirmed by ${\beta}-galactosidase$ activity and Western blot analysis of sfs1-lacZ fusion gene. Transcriptional regulation by cAMP-CRP was also confirmed by Northern blot analysis. End-labelled DNA of the DNA fragment in sfs1 regulation region were used for gel retardation assay to examine the CRP-DNA complex in the presence of cAMP. Results here indicate that CRP binding site in the regulatory region of sfs1 gene is positive regulator for the expression of sfs1 gene.

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Synergistic effects of oxytetracycline and tiamulin against swine respiratory bacteria and their pharmacokinetics in the pigs (돼지호흡기세균에 대한 oxytetracycline과 tiamulin의 시험관내 상승작용 및 돼지에서의 약물동태학)

  • Park, Seung-chun;Yun, Hyo-in;Lee, Keun-woo
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.45-54
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    • 2002
  • The study was carried out to characterize the pharmacokinetics after intravenous (iv, 20 mg/kg) and oral (p.o. 100 mg/kg) administration as oxytetracycline (OTC) and tiamulin (TIA) mixture in swine and to determine interaction between OTC and TIA against various pig pathogenic bacteria. The antibacterial effects of OTC in combination with TIA in vitro showed synergistic effect against Salmonella typhimurium 1925, Pasteurella multocida Type A, P. multocida Type D, Krebsiella Pneumoniae 2001, K. Pneumoniae 1560, K. Pneumoniae 2208, Haemophillus pleuropneumonia S 2, and H. pleuropneumonia S 5, but against additive effect E. coli K88ab and S. choleraesuis on the basis of fractional inhibitory concentration (FIC) index. On the while, after i.v. and p.o. administration of OTC and TIA mixture, each OTC and TIA concentrations in plasma were fitted to an open two-compartment model. After i.v. administration of OTC-TIA mixture, the mean distribution half-life ($T_{1/2{\alpha}}$) of OTC and TIA in plasma showed 0.29 h and 0.17 h, and the mean elimination half-life ($T_{1/2{\beta}}$) of those was 4.36 h and 6.64 h, respectively. The mean volume of distribution at steady state ($Vd_{ss}$) of OTC and TIA was $0.85{\ell}/kg$ and $2.44{\ell}/kg$, respectively. After oral administration of OTC and TIA mixture, the mean maximal absorption concentrations ($C_{max}$) of OTC and TIA were $0.60{\mu}g/m{\ell}$ at 1.07 h ($T_{max}$) and $1.68{\mu}g/m{\ell}$ at 1.85 h ($T_{max}$), respectively. The mean elimination half-life ($T_{1/2{\beta}}$) of those showed 6.84 h and 6.36 h. In conclusion, we could suggest in this study that the combination of OTC and TIA may be recommended for the antibacterial therapy against polymicrobial infections, and both OTC and TIA showed large distribution to tissues and high $C_{max}$ after p.o. administration.

Bacillus subtilis를 이용한 대두 발효식품의 혈전용해능

  • Jeong, Yeong-Gi
    • Proceedings of the Korean Society of Life Science Conference
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    • 2001.06a
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    • pp.67-86
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    • 2001
  • A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

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A Case Study using a Strategic Grid for Effective Cluster Development : Chungbuk Software industry Case (효과적인 클러스터 구축을 위한 전략격자모형 설계 및 사례연구 : 충북SW산업을 중심으로)

  • Hwang, Yoon-Jeong;Kim, Jong-Tae;Kwon, Seong-Taek;Yeon, Seung-Jun
    • Proceedings of the Korea Society of Information Technology Applications Conference
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    • 2006.06a
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    • pp.185-207
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    • 2006
  • 산업단지는 지난 30년간 한국 산업의 성장을 이끌어 온 발전모형으로서 존재하여 왔으나, 최근 지식에 기초한 혁신창출형 경제체제가 국가 및 지역사회의 경쟁력을 위한 핵심요소로 부각되면서 '효율성' 측면에서 그 의미가 크게 퇴색되었다. 이를 위한 대안으로서 '클러스터'가 대두되어 다양한 분석연구가 수행되고 있으며, 정부와 지방자치단체들은 이를 바탕으로 각자의 특색에 맞는 클러스터 조성 정책을 펼치고 있다. 그러나, 기존의 연구들은 클러스터의 종류 및 발전단계에 관한 프레임워크 제시 등의 이론적 수준에 국한되어 있거나, 지역사례 연구를 통한 성공요인분석(CFS) 및 단순한 정책방향 제시 수준에 머물러 있는 한계를 보이고 있다. 본 연구는 '클러스터'에 관한 선행연구를 분석해 보고, 클러스터의 중요한 판단기준이 되는 군집도와 네트워크 연계 정도를 기준으로 한 '$2{\times}2$ 클러스터 전략격자모형'을 효과적인 클러스터 구축전략 수립을 위한 이론적 틀로서 제시하였다. 또한, 분석틀에 실질적인 사례로서 '충북지역의 SW산업'을 전략격자모형에 대응시켜 분석함으로써 전략격자의 유용성을 제시하였다. 이를 위해, 충북지역의 SW 공급업체와 수요업체를 대상으로 설문조사를 실시, 분석한 후 그 결과를 전략격자모형에 대응시켰다. 그 결과, 충북지역의 SW산업은 아직 산업단지 수준에 있는 것으로 분석되었고 충북의 SW산업의 충북 내의 수요만으로는 더 큰 성장이 어려운 것으로 분석, 지역 내에서의 수요창출을 목표로 하는 '단일 클러스터' 구축보다는 지역적 제약을 벗어난 '매가 클러스터'의 구축으로 지역 내외에서의 수요창출이 가능한 클러스터의 구축을 그 대안으로 제시하였다.${\alpha}$에 E. coli Jm109의 plasmid pBX19, pBR322를 전이시켰다. 6. L. lactis ssp. lactis 균주에 lysozyme 처리시 30${\sim}$80%의 생존율을 보였으며, 대부분의 L. acidophilus 균주의 경우 약 70%의 생존율을 보였다. L. casei 102S의 경우는 45분간 처리 시에도 100%의 생존율을 보였다. 8. L. lactis ssp. lactis 균주에 pLZ12를 6.0kV에서 전이시킨 결과 12.5kV에서보다 형질전환 효율이 훨씬 높았으며 lysozyme 처리에 의해 형질전환 효율이 증가되었다. 9. L. acidophilus 균주에 pLZ12를 전이시 6.0kV에서는 전이가 모두 이루어졌으나, 12.5kV에서는 L. acidophilus WIESBY와 NCFM에서 전이가 이루어지지 않았으며, lysozyme 처리 후 pLZ12를 전이시켰을 때 12kV보다 6.0kV에서 형질전환 효율이 증가되었다. 10. Gene Pulser와 Progenitor II를 사용하여 pLZ12를 L. lactis ssp. lactis 균주에 전이하였을 때 Gene Pulser에 비해 Progenitor II의 형질전환 효율이 현저히 떨어졌다. L. acidophilus HY7008과 HY7001은 두 기기 모두 형질전환이 이루어졌으나, L. acidophilus WEISBY와 NCFM은 Progeni-tor II에서 전이가 일어나지 않았으며, Gene Pulser에서 전이균주를 얻어 두 electroporator간에 형질전환 효율의 차이를 보였다. 11. L. casei 102S에 pLZ12

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