• Title/Summary/Keyword: Agar diffusion

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Selection and Antifungal Activity of Antagonistic Bacterium Bacillus subtilis KMU-13 against Cucumber scab, Cladosporium cucumerinum KACC 40576 (검은별무늬병균 Cladosporium cucumerinum KACC 40576에 대한 길항균주 Bacillus subtilis KMU-13의 선발 및 항진균 활성)

  • Park Sung-Min;Lee Jun-Seuk;Park Chi-Duck;Lee Jung-Hun;Jung Hyuck-Jun;Yu Tae-Shick
    • KSBB Journal
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    • v.21 no.1 s.96
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    • pp.42-48
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    • 2006
  • Bacillus subtilis KMU-13 was isolated from the Lillehammer forest soils at Norway and shown a strong antifungal activity on cucumber scab, Cladosporium cucumerinum KACC 40576. B. subtilis KMU-13 produced a maximum level of antifungal substance under incubation aerobically at $30^{\circ}C$, 180 rpm for 48 hours in LB broth containing 0.5% maltose and 0.5% bactopeptone and initial pH adjusted to 6.0. Butanol extract of cultured broth was confirmed inhibitory zone by plate assay and Rf 0.64 value substance by thin layer chromatography (TLC) represented high antifungal activity against C. cucumerinum KACC 40576 and also shown fungal growth inhibitory activity against Botytis cinerea KACC 40573, C. gloeosporioides KACC 40804, D. byoniae KACC 40669, F. oxysporum KACC 40037, F. oxysporum KACC 40052, F. oxysporum f. sp. radicis-lycopersici KACC 40537, F. oxysporum KACC 40902, M. cannonballus KACC 40940, P. cambivora KACC 40160, R. soiani AG-1 KACC 40101, R. solani AG-4 KACC 40142, and S. scleotiorum KACC by agar diffusion method.

Salmonella typhimurium LPS Confers Its Resistance to Antibacterial Agents of Baicalin of Scutellaria baicalensis George and Novobiocin: Complementation of the rfaE Gene Required for ADP-L-glycero-D-manno-heptose Biosynthesis of Lipopolysaccharide

  • Chung, Tae-Wook;Jin, Un-Ho;Kim, Cheorl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.564-570
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    • 2003
  • The antibacterial mechanism of enterobacter Salmonella typhimurium was studied. The rfa (Waa) gene cluster of S. typhimurium encodes the core oligosaccharide biosynthesis of lipopolysaccharide (LPS). Among the rfa gene cluster, we recently cloned the rfaE gene, which is involved in ADP-L-glycero-D-manno-heptose biosynthesis. The rfaE mutant synthesizes heptose-deficient LPS, which consists of only lipid A and 3-deoxy-D-manno-octulosonic acid (KDO), thus making an incomplete LPS and a rough phenotype mutant. S. typhimurium deep-rough mutants with the heptose region of the inner core show a reduced growth rate, sensitivity to high temperature, and hypersensitivity to hydrophobic antibiotics such as baicalin isolated from the medicinal herb of Scutellaria baicalensis Georgi. Thus, in this study, the cloned rfaE gene was added to the S. typhimurium rfaE mutant strain SL1102 (rfaE543), which makes heptose-deficient LPS and has a deep-rough phenotype. The complementation created a smooth phenotype in the SL1102 strain. The sensitivity of SL1102 to bacteriophages was also recovered to that of wild-type strain, indicating that LPS is used as the receptor for bacteriophage infection. The permeability barrier of SL1102 to hydrophobic antibiotics such as novobiocin and baicalin was restored to that of the wild-type, suggesting that antibiotic resistance of the wild-type strain is highly correlated with their LPS. Through an agar diffusion assay, the growth-inhibition activity of baicalin was fully observed in the mutant SL1102 strain. However, only a half of the inhibitory activity was detected in the rfaE complemented SL1102 strain. Furthermore, the LPS produced by the rfaE-complemented SL1102 strain was indistinguishable from LPS biosynthesis of smooth strains.

Efficacy of Antagonistic Bacteria for Biological Control of Rhizoctonia Blight (Large patch) on Zoysiagrass (잔디 갈색퍼짐병(Large patch)의 생물학적 방제를 위한 길항 미생물의 선발과 효력 검정)

  • Jung, Woo-Chul;Shin, Taek-Su;Kim, Bong-Su;Im, Jae-Seong;Lee, Jae-Ho;Kim, Jin-Won
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.43-50
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    • 2008
  • Rhizoctonia blight (large patch) caused by Rhizoctonia solani AG2-2 is one of the major diseases on zoysiagrass in golf courses. In this study, anatgonistic bacteria to R. solani AG2-2 were selected in vitro tests using confrontation bioassay and triple layer agar diffusion method. The most active bacteria, Bacillus subtilis CJ-9 were tested for controlling large patch in pots. Relative Performance Indies (RPI) was used as a criterion for the selection of potential biocontrol agent. B. subtilis CJ-9 showed resistance to major synthetic agrochemicals used in golf course. In field tests at golf course, B. subtilis CJ-9 was more effective in suppression of large patch severity and population development of R. solani AG2-2 in soil than chemical fungicides. B. subtilis CJ-9 could be an alternative to chemical fungicides for eco-friendly management of large patch on zoysiagrass.

Isolation and Optimal Culture Conditions of Brevibacillus sp. KMU-391 against Black Root Pathogens Caused by Didymella bryoniae (덩굴마름병원균인 Didymella bryoniae의 생물학적 방제를 위한 길항세균의 분리 및 특성)

  • Park Sung-Min;Jung Hyuck-Jun;Kim Hyun-Soo;Yu Tae-Shick
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.135-141
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    • 2006
  • We isolated a bacterium which produces antifungal substances from the Sanktpeterburg soils at Russia. The iso-lated strain was identified as Brevibacillus sp. and shown a strong antifungal activity on plant pathogenic fungi. Brevibacillus sp. KMU-391 produced maximum level of antifungal substances under incubation aerobically at $30^{\circ}C$ for 48 hours in trypticase soybean broth containing 1.0% sucrose and 1.0% polypeptone at 180 rpm and initiated pH adjusted to 7.0. Precipitate of culture broth by $30{\sim}60%$ ammonium sulfate precipitation exhibited strong antifungal activity against Didymella bryoniae by dry cell weight. Butanol extract of cultured broth also shown fungal growth inhibitory activity against Botrytis cinerea KACC 40573, Botrytis fabae KACC 40962, Colletotrichum gloeosporioides KACC 40804, Colletotrichum orbiculare KACC 40808, Didymella bryoniae KACC 40669, Fusarium graminearum KACC 41040, Fusarium oxysporum KACC 40037, Fusarium oxysporum KACC 40052, Fusarium oxysporum f, sp. radicis-Iycopersici KACC 40537, Fusarium oxysporum KACC 40902, Monosporascus cannonballus KACC 40940, Phytophthora camvibora KACC 40160, Rhizoctonia solani AG-1(IA) KACC 40101, Rhizoctonia solani AG-4 KACC 40142, and Scleotinia scleotiorum KACC 41065 by agar diffusion method.

Application Potential of Hurdle Technology by Combination of Bacteriocin Produced by Lactobacillus brevis DK25 and Potassium Benzoate (Lactobacillus brevis DK25의 박테리오신과 안식향산칼륨과의 혼용에 의한 Hurdle Technology 적용 가능성)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.364-374
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    • 2011
  • Lactobacillus brevis DK25 isolated from Dongchimi was identified by physiological and biochemical tests and 16S rDNA sequence analysis. Bacteriocin of L. brevis DK25 exhibits inhibitory activity against Enterococcus faecalis and Listeria monocytogenes when using agar well diffusion method. Maximal production of bacteriocin was reached in the beginning of the stationary phase, and inhibitory activity declined after the late stationary phase. This result suggested that bacteriocin was produced in a growth-associated manner. Complete inactivation of bacteriocin activity was observed after treatment with protease, but the activity was stable between pH 4-9 and heat resistant (30 min at $100^{\circ}C$). Bacteriocin showed a concentration-dependent antimicrobial activity against L. monocytogenes KCTC 3569. Moreover, the application experiment showed that combination of bacteriocin (320 AU/ml) with potassium benzoate (0.05%) could significantly reduce the counts of L. monocytogenes KCTC 3569 in mayonnaise during storage at 4 or $25^{\circ}C$ for 10 days. Thus, bacteriocin from L. brevis DK25 may be used for hurdle technology by combination with potassium benzoate in order to increase pathogenic bacteria inactivation in food processing and food safety control.

Bioequivalence of Two Clarithromycin Tablets (클래리스로마애신 정제의 생물학적 동등성 평가)

  • 김종국;이사원;최하곤;고종호;이미경;김인숙
    • Biomolecules & Therapeutics
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    • v.6 no.2
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    • pp.219-224
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    • 1998
  • The bioequivalence of two clarithromvcin products was evaluated with 16 normal male volunteers (age 23-28 yr, body weight 57.5-75.517g) following single oral dose. Test product was ReYon Clarithromycin tablets (ReYon Pharm. Corp., Korea) and reference product was Klarici $d_{R}$ tablets (Abbott Korea). Both products contain 250 mg of clarithromucin. One tablet of the test or the reference product was administered to the volunteers, respectively, by randomized two period cross-over study (2$\times$2 Latin square method). The determination of clarithromycin was accomplished using a modified agar well diffusion bioassay. As a result of the assay validation, the quantification of clarithromycin in human serum by this technique was possible down to 0.03$\mu$g/ml using 100$\mu$l of serum. The coefficient of variation (C.V.) was less than 10%. Average drug concentrations at each sampling time and pharmacokinetic parameters calculated were not significantly different between two products P>0.05); the area under the curve to last sampling time (24 hr) (AU $Co_{24hr}$ (8.10$\pm$ 1.26 vs 8.22$\pm$ 1.627g . hr/ml), AUC from time zero to infinite (AU $Co_{\infty}$) (8.61 $\pm$ 1.28 vs 8.84$\pm$ 1.71 $\mu$g . hr/ml), maximum plasma concentration ( $C_{msx}$) (0.87$\pm$0.22 vs 0.88$\pm$0.19 $\mu$g/ml) and time to maximum plasma concentration ( $T_{max}$) (2.69 $\pm$0.48 vs 2.56$\pm$ 0.51 hr). The differences of mean AU $Co_{24h}$, $C_{msx}$ and $T_{msx}$ between the two products (1.44, 1.39, and 4.65%, respectively) were less than 20%. The power (1-$\beta$) and treatment difference ($\Delta$) for AU $Co_{24hr}$, and $C_{max}$ were more than 0.8 and less than 0.2, respectivly. Although the power for $T_{max}$ was under 0.8, $T_{max}$. of the two products was not significantly different each other (p>0.05). These results suggest that the bioavailability of ReYon Clarithromycin tablets is not significantly different from that of Klarici $d_{R}$ tablets. Therefore, two products are bioequivalent based on the current results. results.sults.sults.s.s.s.s.s.s.s.

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Antimicrobial Activity of Maesil(Prunus mume) Juice against Selected Pathogenic Microorganisms (매실(Prunus mume) 착즙액의 식중독 유발균에 대한 항균 작용)

  • 이현애;남은숙;박신인
    • The Korean Journal of Food And Nutrition
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    • v.16 no.1
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    • pp.29-34
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    • 2003
  • Prunus mume has been used for the folk medicine by many old civilizations to treat food-borne diseases or enteric disorders. This study was carried out to examine the antimicrobial effect of juice from Prunus mume against pathogens such as Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Listeria monocytogenes and Bacillus cereus. The juice of Prunus mume had the strongest antimicrobial activity to Sal. enteritidis. The concentrations of Prunus mume juice for the formation of clear zone were 1% for Sai. enteritidis(15.0mm), 3% for Lis. monocytogenes(14.7mm), and 5% for Bac. cereus(14.75mm), Esc. coli(13.45mm) and Sta. aureux(11.9mm). The growth of all tested microorganisms was inhibited apparently in tryptic soy broth containing 3% and 4% Prunus mume juice. And it was found that the Prunus mume juice showed the highest antimicrobial properties, followed by Sal. enteritidis, Bac. cereus, Sta. aureus, Lis. monocytogenes, Esc. coli.

The Production of Traditional Alcoholic Beverage in Containing Medicinal Herb (한약재를 첨가한 전통주개발에 관한 연구)

  • Kim, Young-Sook;Park, Young-Sook
    • Food Science and Industry
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    • v.40 no.2
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    • pp.83-89
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    • 2007
  • 저온발효와 고온발효에 의하여 만들어진 민속주를 증류하여 알콜돗를 45%로 같이 맞추어 성분함량을 비교하였다. 발효온도에 따른 환원당의 함량을 나타내고 있으며 저온발효 민속주는 15 mg/100 mL, 고온발효 민속주는 5 mg/100 mL로 나타났다. 즉 저온 발효에서 환원당의 양이 많았으며, 그러나 시중에 판매되고 있는 소주보다는 낮은 경향을 보였다. 향기 성분에 생성과정에 영향을 주는 Fusel alcohol을 볼 때 반적으로는 발효액에 아미노산이 없는 것보다는 많을 수록 많은 양의 fusel alcohol을 생성되기는 하지만, fusel alcohol을 생성하기위한 조건은 효모의 종류, 발효조건, 발효온도, 발효액의 조성 등의 복잡한 인자가 관련된다. 본 연구에 제조한 발효주를 소주의 아미노산과 비교할 때 아미노산 함량이 전반적으로 20-100도 낮은 함량치로 나타났다. 한약재 첨가로 인하여 면역증강, 살세포 반응 억제효과, 기능성 전통주로 좋은 발효주로 인정되지만 한약재특유의 강한 향기를 약화시키는 연구가 금후에 보완되어야 한다고 본다. 발효온도에 따른 아미노산의 함량은 저온발효 조미주에서 cysteine, valine, mrthionine, isoleucine, phenylalanine이 많았으며, 고온발효 조미주에서는 serine, glycine, leucine이 많았다. 한 alanine, tyrisine, lysine은 고온발효주에는 있으나, 저온발효주에서는 나타나지 않았다. 그러나 저온발효주와 고온 발효주 모두 시중에 판매되고 있는 소주보다는 아주 낮은 경향을 보였다. 저온발효와 고온발효에 의하여 만들어진 민속주를 증류하여 알콜 도수를 45%로 같이 맞추어 성분함량을 비교하였다. 발효온도에 따른 환원당의 함량을 나타내고 있으며 저온발효 민속주는 15 mg/100 mL, 고온발효 민속주는 5 mg/100 mL로 나타났다. 즉 저온 발효에서 환원당의 양이 많았으며, 그러나 시중에 판매되고 있는 소주보다는 낮은 경향을 보였다. 아미노산 실험에서는 소주와 45% 고온 발효주를 비교 할 때 serine, glycine, cysteine, methionine, phenylalanine 등은 100배, alanine, valine, isoleucine 80배, ammonia 20배 낮은 수치를 나타냈다. OD 측정의 실험결과 또한 control과 비교시 40% 발효주에서는 10배, 45% 고은 발효주에서는 100배 낮은 측정치가 나타났으므로 한약재의 색소성분을 휘발시키는 기술이 요구된다고 본다. 항균성실험에서 항균성 측정은 06 cm paper disk agar diffusion법을 이용하였으며, 43%의 발효주와, 45% 고온 발효주가 항균력이 가장 강력한 0.95 cm의 영향을 나타냈다. 사용한 사용한 Gram 양성, Gram 음성 균주는 Escherichia coli KCCM 11591를 제외하고는 0.8 - 0.95 cm로 항균력이 강했으며, Gram negitive의 Pseudomonas aeruginosa KCTC 1750 에서는 43% 발효주에는 0.95 cm, 45% 고은 발효주에는 0.95 cm의 항균성을 나타냈으며 관능평가에서도 가장 높게 났다. 관능평가에서는 45% 고온 발효주가 가장 높게 나타났으며, 항산화성 실험에 나타난 저온 45%의 갈색도의 측정과는 항산화성에서는 좀 다른 결과를 나타낸다. 그러나 항균성이 가장 높게 나타난 43-45%와 관능평가에서 가장 높게 나타난 45% 고온 발효주를 볼 때 본 연구에서는 고온 발효주 45%가 가장 우수한 전통주로 조사되었다. 발효주를 소주의 아미노산과 비교할 때 아미노산 함량이 전반적으로 100배 정도 낮은 함량치로 나타났으므로, 한약재 첨가로 인하여 면역증강, 살세포반응 억제효과, 기능성전통주로 좋은 발효주로 인정되지만 한약재특유의 강한 향기를 약화시켜서 부드럽고 은은한 전통발효주의 연구가 금후에 보완되어야 한다고 본다.

Antimicrobial Activity Screening of Sixty-four Evergreen Woody Species According to Extraction Conditions against Trichophyton mentagrophytes (상록성 목본 64종의 추출조건에 따른 무좀원인균의 항균활성 스크리닝)

  • Jang, Bo Kook;Chi, Lai Won;Cho, Ju Sung;Lee, Cheol Hee
    • Korean Journal of Plant Resources
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    • v.31 no.4
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    • pp.330-341
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    • 2018
  • This study was performed to investigate and measure the antimicrobial activity of evergreen woody species extracts on Trichophyton mentagrophytes. To do this, leaves and stems were collected from Wando and Jeju islands, and were used for the extraction with different solvents (i.e., distilled water, 80% ethanol, and 100% methanol), and at different ultrasonic extracting times (i.e., 15, 30, and 45 minutes). The experiment was conducted by using the agar diffusion method. The clear zone was measured after incubating the paper disc containing the plant extract in a bacterial culture medium. The controls were synthetic antimicrobials, methylparaben and phenoxyethanol, at concentrations of 0.4, 1, 2, and 4 mg/disc. Altogether, extracts of 56 out of 64 species used in this study had inhibitory activity, which confirmed their antimicrobial activity against Athlete's foot. Among them, the crude ethanolic extract of Elaeocarpus sylvestris in 45 min showed a zone of inhibition < 20.2 mm, while the clear zone of Actinodaphne lancifolia ethanolic extraction for 30 min was 23.5 mm. Also, Quercus acuta, Dendropanax morbiferus and Daphne odora showed clear zones of 28.0 mm (45 minutes ethanolic extraction), 20.5 mm (45 minutes crude methanolic extraction) and 19.7 mm (45 minutes methanolic extraction), respectively. Thus, these results confirm that the extracts of evergreen woody species have therapeutic potential against Athlete's foot, and suggest that in order to extract adequate amounts of antimicrobial substance from the plant sources, ideal extraction condition has to be considered.

A Study of Antibiotic Resistance of Enterococci Isolated from Recycling Water of Floor Fountains in Gwangju Area (광주지역내 바닥분수 재이용수에서 분리한 장구균의 항생제 내성 연구)

  • Kim, Yeon-Hee;Song, Hyeong-myeong;Choi, Yeong-Seop;Lee, Yoon-Kook;Park, Jin-Hwan;Kim, Sun-Jung;Kim, Ha-Ram;Kang, Yu-Mi;Bae, Seok-Jin;Cho, Yong-Gwan
    • Journal of Environmental Health Sciences
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    • v.45 no.3
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    • pp.222-230
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    • 2019
  • Objectives: This study aims to evaluate water quality in terms of microorganisms and identify the antibiotic resistance of Enterococci isolated from the recycling water in floor fountains at three parks and one reservoir in the Gwangju area. Methods: Water samples were analyzed for Enterococci using membrane Enterococcus indoxyl ${\beta}$ d glucoside agar (mEI) as described in USEPA Method 1600. The vancomycin-resistant Enterococci with VanA and VanB were identified by PCR. An examination of the antibiotic resistance of isolates against 14 antibiotics was performed by the disk diffusion method. Results: The drinking water quality criterion was exceeded for total colony counts in 68% of all recycling water samples. The average concentration of total califorms and fecal coliforms was 139,325 and 413 CFU/100 mL, respectively. VanA and VanB were not detected from the isolates. We found the antibiotic resistant Enterococci strains to be E. faecalis, E. faecium, E. durans, E. mundtii, E. hirae, and E. thailandicus. The isolates were resistant to Rifampin (50%), Erythromycin (25.8%), Tetracycline (10.2%), Nitrofurantoin (8.1%), Minocycline (3.1%), Erythromycin (1.2%), Penicillin (0.7%), Norfloxacin (0.5%), and Teicoplanin (0.5%) among the 14 antibiotics tested. Antibiotic resistance tests for Enterococci from the recycling water of floor fountains resulted in 30.2% showing resistance to two or more antibiotics. Conclusions: These results showed that the multi-antibiotic resistance of Enterococci, E. coli, and others should be investigated continuously in each environment field.