• Title/Summary/Keyword: resistance bacteria

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Transfer of R plasmids of Bacterial Isolates and Their Cloned R Genes in Natural Wastewater Environments (I) -Cloning of $Km^rCm^r$Gene- (하폐수의 자연환경에서 R plasmid와 재조합 유전자의 전이특성( I ) -$Km^rCm^r$유전자의 클로닝-)

  • 김치경;이성기
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.447-453
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    • 1989
  • In order to study the transfer of antibiotics resistance genes of the genetically cloned bacteria in water environments, DK1 strain, which is resistant to kanamycin (Km), chloramphenicol (Cm), streptomycin (Sm), and sulfadiazine (Su), was selected from the Gram-negative bacterial isolates from wastewater. One of 4 plasmids harboured in the DK1 strain was found to possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and be about 68 kb in size, and it was designated as pDK101. The plasmid of pDK101 was also found to have 16, 32, and 6 restriction sites for EcoRI. .PstI, and SalI, respectively. From the digestion fragments of pDK101 plasmid and pKT230 used as a vector by EcoRI restriction endonuclease, pDT309 and pDT529 were constructed as chimeric plasmids which possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and are 30.9 and 52.9 kb in size, respectively. When the chimeric plasmids were trasformed into E. coli C600 or E. coli HB101, transformants of DKC601, DKC602, DKH102, and DKH103 were obtained as cloned bacterial cells. The Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ genes were well expressed in those cloned cells and the chimeric plasmids were clearly detected in the cloned cells of DKC601 and DKH103.

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Selection of the Auxin and ACC Deaminase Producing Plant Growth Promoting Rhizobacteria from the Coastal Sand Dune Plants (Auxin과 ACC Deaminase를 생산하는 사구식물 복원용 근권세균의 선발)

  • Lim, Jong-Hui;Kim, Jong-Guk;Kim, Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.268-275
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    • 2008
  • In order to develop the multi-functional rhizobacteria that can exert positive effect on the growth of plants growing in the coastal sand dune located along East Coast of Korea, rhizospheral bacteria of 11 different plants from this area were isolated 1,330 rhizobacteria. Among these, 23 strains were able to produce auxin and had spectrum of antagonism toward various phytopathogenic microbes. To know the mechanism of this antifungal activity, these 23 strains were subjected to further analyses; 19 strains of these produced siderophore as determined by color reaction on CAS-blue plate, 4 strains produced antifungal cellulase as judged by color change on CMC-Congo red plate, 17 strains were able to utilized insoluble phosphate salts, also determined by clear zone formation on PVK medium. Identification of the strain was assigned to all 23 strains by l6s rDNA sequence analysed, and all were identified to be in the genus of Bacillus and Pseudomonas. One strain of these, denoted Pseudomonas fluorescens IB4-14, showed ACC deaminase activity which is known to be involved in the resistance of environmental stress such as salt and drought. Also, P. fluorescens IB4-l4 showed the germination stimulation and roots growth promoting activity on the in vivo assay of Lysimachia mauritiana Lam. (spoonleaf yellow loosestrife).

Cholesterol Lowering Effect of Lactobacillus plantarum Isolated from Human Feces

  • Ha Chul-Gyu;Cho Jin-Kook;Lee Chi-Ho;Chai Young-Gyu;Ha Young-Ae;Shin Shang-Hun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1201-1209
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    • 2006
  • The purpose of this study was to isolate probiotic lactic acid bacteria (LAB) that produce bile salt hydrolase (BSH), and to evaluate its effects on serum cholesterol level. One-hundred-twenty bacterial colonies were initially isolated from human feces, and five strains were selected after screening based on their resistance to acids, tolerance against bile salts, and inhibitory activity on Escherichia coli. The Lactobacillus plantarum strain with the highest level of BSH activity was identified using 16S rRNA sequences, and was named L. plantarum CK 102. L. plantarum CK 102 at a level of 1.36$\times$10$^8$cfu/ml survived in pH 2 buffer for 6 h and exhibited excellent tolerance for bile salt. Coculturing the strain with E. coli in MRS broth resulted in strong inhibition against growth of E. coli at 18 h. Furthermore, the potential effect of CK 102 on serum cholesterol level was evaluated in rats. Thirty-two rats [Sprague-Dawley (SD) male, 129$\pm$l g, 5 weeks old] were divided into four groups of eight each. For six weeks, Group 1 was fed a normal diet (negative control); Group 2 was fed a cholesterol-enriched diet (positive control); Group 3 was fed a cholesterol-enriched diet plus L. plantarum CK 102 at 1.0$\times$10$^7$cfu/ml; and Group 4 was fed a cholesterol-enriched diet plus L. plantarum CK 102 at 5.0$\times$10$^7$cfu/ml. Blood samples were collected, serum lipids were analyzed, and weights of the organs were measured. Total blood cholesterol level, triglyceride, LDL-cholesterol, and free-cholesterol values were lower in rats that were fed 1. plantarum CK 102 than in those not fed L. plantarum CK 102. This cholesterol lowering effect implies that L. plantarum CK 102 could be utilized as an additive for health-assistance foods. In conclusion, these results suggest that the 1. plantarum CK 102 isolated could be used commercially as a probiotic.

Evaluation of the Antimicrobial Activities of 35 Seaweed Extracts against Pathogenic Bacteria and Candida sp. (35종 해조류 추출물의 병원성 세균 및 Candida sp. 진균에 대한 항균 활성 평가)

  • Kim, Mi-Sun;Kwon, Kyung-Jin;Lee, Min-Jin;Ahn, Seon-Mi;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.144-151
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    • 2012
  • In the course of this study aimed at the development of functional food ingredients from seaweeds, the in vitro antimicrobial activities of methanol extracts prepared from 35 different seaweeds (17 phaeophyta, 11 rhodophyta and 7 chlorophyta) were determined against food-borne diseases and pathogenic microorganisms including multi-drug resistant (MDR) Pseudomonas sp. and Candida sp. Based on disc-diffusion assays at 500 g/disc concentration of the methanol extracts, Ishige okamurai, I. foliacea, Sargassum confusum, and S. yamade exhibited strong antibacterial activities in a broad-spectrum, except against Pseudomonas aeruginosa. In addition to the latter four seaweeds, Ecklonia stolonifera, E. cava and Eisenia bicyclis also demonstrated antifungal activity against C. albicans. Among these 8 selected seaweeds, I. okamurai, I. foliacea, and S. yamade exhibited strong hemolytic activity (55-93%) at 500 g/ml against human RBC. Organic solvent sequential fractions using hexane, ethylacetate and butanol, and water residues were prepared from the 8 selected seaweeds and their anti-Candida sp. activities were further determined. The ethylacetate and butanol fraction of I. okamurai, and the hexane fraction of I. foliacea demonstrated antifungal activity against MDR-pathogenic Candida sp. Although the solvent fractions had no activity against MDR-Pseudomonas sp., our results suggest that seaweeds, especially Ishige okamurai, I. foliacea, S. confusum, and S. yamade could be developed as broad-spectrum antimicrobial ingredients.

Anti-bacterial Effect of Psidium guajava and Geranium thunbergii Extracts on Listeria sp. Isolated from Fishery Products (수산가공품에서 분리한 Listeria sp.에 대한 구아바(Psidium guajava)잎과 현초(Geranium thunbergii) 추출물의 항균활성)

  • Kim, Yang-Ho;Kim, Tae-Yong;Kim, Jin-Su;Choi, Jae-Woo;Lee, Su-Jeong;Cha, So-Young;Shin, So-Dam;Jeon, Mi Hyeon;Noh, Da-In;Lee, Eun-Woo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.2
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    • pp.237-243
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    • 2020
  • Listeria sp. is one of the pathogenic bacteria causes the infection listeriosis, through mainly raw food such as fishery food, dairy food and vegetables. Listeria sp. is a Gram-positive, non-spore-forming, motile, and facultative anaerobic bacterium. Because of the tolerance of Listeria sp. to low temperature and high salt concentration, it is very difficult to prevent them contaminated in the food, which do not require heating, especially, such as raw fishery products. So prevention and removal of bacterial contamination at the food manufacturing stage is the best method. In this study, therefore, several natural products including Psidium guajava and Geranium thunbergii were screened to investigate the antibacterial activity against Listeria sp., with expectation of fewer side effects and fewer resistance problems. Significant effects of two extracts were confirmed by well diffusion assay, MIC assay, and growth inhibition assay. P. guajava and G. thunbergii showed MIC values at 64-256 ㎍/mL meaning strong antibacterial activities against 6 kind of Listeria sp. tested. And the growth of Listeria sp. in the liquid media was actually inhibited by the addition of these two extracts.

Effect of Bacillus subtilis C4 and B. cereus D8 on Plant Growth of Canola and Controlling Activity Against Soft Rot and Stem Rot (Bacillus subtilis C4와 B. cereus D8에 의한 유채의 생육증대 및 무름병과 균핵병 방제효과)

  • Lee, Jae-Eun;Lee, Seo-Hyeun;Park, Kyung-Soo;Park, Jin-Woo;Park, Kyung-Seok
    • The Korean Journal of Pesticide Science
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    • v.13 no.4
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    • pp.275-282
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    • 2009
  • The effect of two plant growth-promoting rhizobacteria (PGPR) on plant growth and systemic protection against soft rot disease and stem rot disease of canola (Brassica napus), caused by Erwinia carotovora and Sclerotinia sclerotiorum was investigated in a laboratory and a greenhouse. Selected PGPR strains C4 and D8 were treated to canola seeds by soaking. Strains C4 and D8 significantly not only increased plant height and root length about 74% and 40.3% and also reduced disease severity of soft rot disease by 80% by C4 and D8 respectively, compared to the control. Especially strain C4 showed antifungal activity against 6 fungal pathogens, S. sclerotiorum, Rhizoctonia solani, Botrytis cinerea, Fusarium oxysporum, Phytophthora capsici and Colletotrichum acutatum. In greenhouse experiment, the seed treatment of both of them increased plant height, leaf width and leaf length of canola plant to 19.5% and 24.9%, 11.3% and 15.3%, and 14.1% and 20.7% by C4 and D8, respectively, and reduced disease severity of S. sclerotiorium. These results indicate that these two PGPR strains can decrease disease severity and increased plant growth under greenhouse condition. Therefore, these two bacteria have a potential in controlling Sclerotinia stem rot of canola. These strains have to investigate under field condition to determine their role of antibiosis, induced systemic resistance and plant growth promotion on canola.

Antibacterial Activity of Graviola extract to inhibit the Staphylococcus epidermidis (Staphylococcus epidermidis를 억제하는 Graviola 추출물의 항균활성)

  • Choi, Jong-Hwa;Ohk, Seung-Ho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.5
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    • pp.667-673
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    • 2017
  • This study was carried out to verify the antimicrobial effect of Graviola leaves against pathogenic bacteria for the purpose of developing an antibacterial material. The extraction conditions of graviola leaves were $60^{\circ}C$, $80^{\circ}C$ and $98^{\circ}C$, and graviloa leaves werewater extracted at $60^{\circ}C$, $80^{\circ}C$ and $98^{\circ}C$.The extraction yields and extraction conditions were determined. The antimicrobial activity against S. epidermidis, S. aureus and E. coli TOP10 was evaluated by agar diffusion method. The extraction yields were 3.02%, 14.73% and 20.76% at $60^{\circ}C$, $80^{\circ}C$ and $98^{\circ}C$, respectively. The antimicrobial activity against S. epidermidis, S. aureus and E. coli TOP10 was found in S. epidermidis.In the samples extracted at $98^{\circ}C$, a clear zone of 13 mm was observed at 200 mg/mL and of 20 mm at 500 mg/mL. The MIC was 100 mg/mL. The higher the extraction temperature and concentration, the better was the growth inhibition effect. As a result, the natural antimicrobial activity contained in natural materials can solve the problem of resistance to antibiotics. It is considered that antimicrobial activity against S. epidermidis in skin is highly applicable to basic cosmetics and cosmetic materials.

Characteristics of Microorganisms Contaminating Seafood Cooking Drips Exposed to Gamma Irradiation (감마선 조사된 수산 자숙액의 오염 미생물군 특성)

  • Choi, Jong-Il;Kim, Yeon-Joo;Kim, Jae-Hun;Chun, Byung-Soo;Ahn, Dong-Hyun;Kwon, Joong-Ho;Hwang, Young-Jung;Byun, Myung-Woo;Lee, Ju-Woon
    • Food Science and Preservation
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    • v.16 no.2
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    • pp.286-291
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    • 2009
  • Microorganisms in seafood cooking drips were counted and identified. Total viable cell counts were 6.40 and 3.10 log CFU/g in cooking drips of Hizikia fusiformis and Thunnus thynnus, respectively. However, microbial populations fell with increased irradiation doses. In H. fusiformis cooking drips, a 5-log reduction in total aerobic bacteria was obtained by irradiation with 5 kGy. In T. thynnus cooking drips, however, contaminating microorganisms were more resistant to gamma irradiation and only a 1-log reduction was seen. DNA sequence analysis showed that the principal contaminating microorganisms in H. fusiformis and T. thynnus cooking drips were Lactobacillus and Bacillus species, respectively. Therefore, the high irradiation resistance of T. thynnus cooking drips microbes may result from spore formation by Bacillus species.

Insect Resistance of Tobacco Plant Expressing CpBV-ELP1 Derived from a Polydnavirus (폴리드나바이러스 유래 CpBV-ELP1 발현 담배의 내충성)

  • Kim, Eunseong;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.56 no.1
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    • pp.19-28
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    • 2017
  • Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.

Probiotic Effects of Lactobacillus plantarum and Leuconostoc mesenteroides Isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum과 Leuconostoc mesenteroides의 프로바이오틱 효과)

  • Lee, Kyung-Hee;Bong, Yeon-Ju;Lee, Hyun Ah;Kim, Hee-Young;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.1
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    • pp.12-19
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    • 2016
  • The probiotic effects of kimchi lactic acid bacteria (LAB), Lactobacillus plantarum (Lab. plantarum) and Leuconostoc mesenteroides (Leu. mesenteroides), were studied. Lab. plantarum KCCM 11352P (LPpnu) and Leu. mesenteroides KCCM 11353P (LMpnu) were isolated from kimchi and were the predominant LAB. We compared their probiotic effects with Lactobacillus rhamnosus GG (LRgg), a well-known probiotic LAB. LPpnu showed better probiotic activities than LRgg. LMpnu also exhibited almost equal activities as LRgg. These two kimchi LAB strains exhibited resistance to gastric and bile acid, adhesion to intestines, and thermal stability. In particular, LPpnu showed excellent probiotic properties. In addition, LPpnu showed greater antioxidant activity by scavenging DPPH radicals or hydroxyl radicals than LMpnu or LRgg. LPpnu also inhibited growth of HT-29 human colon cancer cells by inducing apoptosis, increasing Bax and suppressing Bcl-2 expression compared to LMpnu or LRgg. Taken together, LPpnu and LMpnu could be used as probiotics, and LPpnu exhibited the most beneficial probiotic activities with anti-oxidant and anti-cancer properties.