• Title/Summary/Keyword: agar diffusion method

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Screening of New Bioactive Materials from Microbial Extracts of Soil Microorganism (I) Antimicrobial Activity from 200 Sampled Using Microdilution Assay

  • Jung, Sang-Oun;Kim, Joon;Chang, Il-Moo;Ryu, Jae-Chun
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.278-285
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    • 1998
  • The microdilution assay recommended by NCCLS (National Committee for Clinical Laboratory Standards) is one of the standardized methods of antibiotic susceptibility test. This method has been widely used clinically to obtain MIC values of antibiotics on pathogenic microorganisms. It is more convenient, rapid and simple to test many samples than other test methods such as agar diffusion assay and broth macrodilution assay. The screening of antimicrobial agents from microbial extracts is too laborious in its process. Therefore, a number of screening methods having more simple procedure have been developed. In our laboratory, we applied microdilution assay for screening the antimicrobial agents. This assay showed dose-response results and was more sensitive than disc diffusion assay in our system. We tested 200 samples of microbial extracts originated from 100 microbial strains and selected several samples as potential candidates. In this report, we show that the microdilution assay is more convenient method in screeing of antibiotic susceptibility than those previously reported.

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Serovars of Xanthomonas campetris pv. oryzae Collected from Korea and Serological Diagnosis of Bacterial leaf Blight (우리나라 벼 흰빛잎마름병균 (Xanthomonas campestris pv. oryzae)의 혈청학적 분류 및 진단)

  • Choi J. E.;Lee D. K.;Seo J. H.
    • Korean Journal Plant Pathology
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    • v.1 no.1
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    • pp.61-66
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    • 1985
  • Seventy-one strains collected from Korea were classified into three serovars (designated A, B-I and B-II) by using agar gel diffusion test with the antisera produced against Xanthomonas campestris pv. oryzae isolates Q7472 and Q7502. Of 71 isolates tested, 65 isolates belonged to serovar A, 5 isolates were serovar B-I, and one isolate was serovar B-II. The isolates of serovar B-I and B-II could be distinguished clearly from those of serovar A showing marked autoagglutination. Xanthomonas campestris pv. oryzae was serologically diagnosed in rice leaves by agar gel diffusion tests, possibly being distinguished from Xanthomonas campestris pv. olyzicola and E. herbicola. The pathogen could be also serologically detected from the extracts of diseased leaves, squeezed immediately, heated at $100^{\circ}C$ or incubated in PSA. Serological detection of Xanthomonas campestris pv. oryzae is a more reliable and less time-comsuming method.

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Acaricidal and antibacterial toxicities of Valeriana officinalis oils obtained by steam distillation extraction (수증기 증류 추출법에 의해 얻어진 미국산 길초근 정유의 살비효과 및 항균효과)

  • Choi, Seon-A;Lee, Hoi-Seon
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.19-23
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    • 2019
  • The chemical compositions of the essential oil of Valeriana officinalis roots obtained by steam distillation method were analyzed by GC-MS. The 16 constituents were identified in the V. officinalis oil, and the most abundant compounds were patchouli alcohol (18.69%) and ${\beta}$-gurjunene (15.26%). Acaricidal effects of the V. officinalis oil were evaluated against Tyrophagus putrescentiae, Haemaphysalis longicornis larva and H. longicornis nymph by contact bioassay. The $LD_{50}$ values against T. putrescentiae, H. longicornis larva and H. longicornis nymph were 28.01, 178.26 and $207.98{\mu}g/cm^2$, respectively. Agar disc diffusion bioassay showed the antibacterial activity of the V. officinalis oil against foodborne pathogens, especially L. monocytogenes. These results showed that the essential oil of V. officinalis roots derived from USA has a potential for development as acaricide and antimicrobial.

Antimicrobial susceptibility features of porcine respiratory bacterial pathogens by modified broth dilution method (돼지 호흡기질병 병인체의 항균제 감수성 조사)

  • 송동준;서동균;이춘식;배영찬;김원일;김봉환
    • Korean Journal of Veterinary Service
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    • v.23 no.1
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    • pp.19-28
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    • 2000
  • There are several main antibacterial susceptibility tests, such as agar dilution method, broth dilution method and disk diffusion technique. Especially, for minimal inhibitory concentration (MIC) test, agar dilution method has been widely used. But that method is so complicated and bothering that it's difficult to treat a large amount of strains. On the other hand, modified broth dilution method(add 1% glucose and 0.018% phenol red as a pH indicator to broth) is fast and easy to perform. Most of all, it can visualize the result by color. The MICs of 22 antibiotics Including penicillins, aminoglycosides, cephalothin, chloramphenicol, lincomycin, ceftiofur, vancomycin and quinolones, erythromycin, colistin. sul-fadimethoxine, trimethoprim for arcanobacterium pyogenes 14 strains, actinobacillus pleuropneu-moniae 41 strains and pasteurella multocida 37 strains, which were collected from porcine during 1996 ∼ 1999, were determined by modified broth dilution method. Actinobacillus pleuropneumoniae was highly susceptible to all kinds of quinolones such as ciprofloxacin, enrofloxacin and norfloxacin and to all aminoglycosides, like gentamicin, apramycin, kanamycin and ampicillin, cephalothin and ceftiofur. But It was quite resistant to solfadimethoxin, colistin and vancomycin. Pasteurella multocida was found to have high susceptibility to ampicillin, cephalothin, chlorampenicol and gentamicin but had mid-degree susceptibility to other aminoglycosides. In addition, it was susceptible to norfloxacin and nalidixic acid, but not to newer fluoroquinolone like ciprofloxacin and enrofloxacin and it was resistant to colistin and kanamycin. Arcanobacterium pyogenes was highly susceptible to most of quinolones such as cipoofloxacin, enrofloxacin and norfloxacin and gentamicin and penicillin G. But it also obtained high resistance against the early quinolone, nalidixic acid and aminoglycosides such as amikacin, apramycin and kanamycin and erythromycin, chlorampenicol, tetracyclin and vancomycin.

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Antimicrobial Efficacy of Fermented Dark Vinegar from Unpolished Rice (현미 발효 흑초의 항균활성)

  • Choi, Hakjoon;Gwak, Gyeongja;Choi, Dabin;Park, Jaeyoung;Cheong, Hyeonsook
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.97-104
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    • 2015
  • Vinegar is a widely used acidic seasoning and can be manufactured using various methods and bases, including cereals, wheat, and fruits. Most studies on vinegar have been conducted to evaluate its antioxidant activity. In the present study, fermented dark vinegar (FDV) produced from unpolished rice was examined for its antimicrobial activity, biochemical content, including the amounts of sugar, total soluble sugar, organic acid, and free amino acids, and pH and physiological activity. The antimicrobial efficiency of FDV was assessed using the paper disc-agar diffusion method. FDV exhibited strong antimicrobial activity against the pathogenic bacteria and yeast strains that were tested. In fact, the activity of FDV was shown to be higher than that of the commercial antibiotics carbenicillin (50 µg/ml) and tetracycline (50 µg/ml) against Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Yersinia enterocolitica, and Lodderomyces elongisporus. The antioxidant activity of FDV and ascorbic acid was evaluated. Using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method, we found that FDV has the highest activity of the antioxidants. After spreading FDV onto tryptic soy broth and yeast extract-peptone-dextrose agar media, the microbial strains were isolated and characterized through physiological and biochemical analysis. Based on 16S ribosomal DNA sequence analysis, the isolated microorganisms exhibited a close similarity to Acetobacter papayae, Acetobacter pasteurianus, and Acetobacter peroxidans.

Microbiological Quantitative Assay of Pivampicillin.HCl (Pivampicillin.HCl에 관(關)한 연구(硏究) I -미생물학적(微生物學的) 정량법(定量法)-)

  • Lee, Han-Pyo;Kim, Chung-Kyu;Sim, Young-Ki;Yong, Kun-Ho
    • Journal of Pharmaceutical Investigation
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    • v.3 no.1_2
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    • pp.16-22
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    • 1973
  • Biological determination of Pivampicillin hydrochloride was examined and an available method has been established. Unhydrolyzed Pivampicillin hydrochloride has no antibacterial activity, but it exhibits an antibacterial activity hydrolyzed with liberation of ampicillin by nonspecific enzymes present in blood and tissues from various species in vitro and in vivo. Using these properties, Pivampicillin hydrochloride can be determined as ampicillin by use of an agar-diffusion method with Bacillus subtilis as the assay organism.

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한국산 도꼬마리 추출물의 항균효과 및 분리 정제

  • 김현수;신재욱
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.183-188
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    • 1997
  • Antimicrobial activity of various extracts of Xanthium strumarium L. was tested against 25 strains of bacteria, yeast and fungus. The crude ethylacetate extract exhibited strong growth inhibition to the tested strains with the exception of partial Gram-negative bacteria. The property of antimicrobial compound was very stable under heat treatment at $120^{\circ}C$, but it was unstable in acid (pH 3.0) and alkali (pH 10.0) treatment. The antimicrobial compounds were purified by boiling water extraction, ethylacetate extraction, charcoal column chromatography, silica gel column chro- matography and reverse phase HPLC. The purified compound A and B were detected in a single peak (each above 98% purity) through the HPLC analysis. The compound A and B showed a strong growth inhibition against Gram-negative and positive bacteria in the agar diffusion method. When tested by the FDA method using the esterase, compound A mainly inhibited the growth of bacteria and compound B showed the growth inhibition of both bacteria and yeasts.

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Antimicrobial Activity of Extracts and Coumaric Acid Isolated from Artemisia princeps var. orientalis (쑥의 추출물 및 Coumaric Acid의 항균활성)

  • 박석규;박종철
    • KSBB Journal
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    • v.9 no.5
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    • pp.506-511
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    • 1994
  • Antimicrobial activity of methanol extract and fraction from mugwort leaves(Artemisia princeps val. orientalis) was investigated for the screening of natural antiwucroblal components. By using agar diffusion method, ethyl acetate(EtOAc) layer fractionated from methanol extract of mugwort leaves showed the highest inhibitory effects against tested microorganisms. The ortho-coumaric acid(200∼600ppm) isolated from EtOAc layer showed strong antibacterial activities for Bacillus subtilis and Salmonella typhimurium. As derivatives of o-coumaric acid, antibacterial activity of para-coumaric acid was 1.2∼1.7 fold higher than that of o-coumaric acid. Three types of coumaric acids strong inhibited the growth of B. subtilis in the culture medium. Growth of S. tyhimurium, P. aeruginosa and S. aureus were effectively inhibited by o-, m- and p-coumaric acids, respectively. Minimum inhibitory dose of p-coumaric acid for B. subtilis was $\100∼200mu\textrm{g}$/paper disk.

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Antimicrobial Efficiency in the Fermented Slurry of Unpolished Rice (현미 발효 슬러리의 항균활성)

  • Choi, Hakjoon;Gwak, Gyeongja;Choi, Dabin;Park, Jaeyoung;Cheong, Hyeonsook
    • Microbiology and Biotechnology Letters
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    • v.43 no.4
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    • pp.307-313
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    • 2015
  • Unpolished rice (UR) is considered to be a healthy alternative to white rice when coping with chronic diseases. In the present study, the fermented slurry of unpolished rice (FSUR) was evaluated with respect to its antimicrobial activities and biochemical characteristics, including the quantities of sugar, total soluble sugar, organic acids, free amino acids, pH, and physiological activity. The antimicrobial efficiency of FSUR was assessed using the paper disc-agar diffusion method. FSUR exhibited strong antimicrobial activity against six pathogenic bacterial strains (Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, and Yersinia enterocolitica) and two fermentation strains (Gluconacetobacter intermedius and Lodderomyces elongisporus). The antimicrobial activity of FSUR was higher than the commercial antibiotics, carbenicillin ($50{\mu}g/ml$) and tetracycline ($50{\mu}g/ml$) against S. aureus, E. coli, L. monocytogenes, P. aeruginosa, S. typhimurium, Y. enterocolitica, and L. elongisporus. Also FSUR had a high antioxidant activity. The microorganisms were isolated from FSUR using tryptic soy broth and yeast extract-peptone-dextrose agar media. The isolated microorganisms were characterized using physiological and biochemical analyses as well as by 16S rRNA gene sequencing and phylogenic analysis. 16S rRNA gene sequence analysis showed that the isolated microorganisms had a high similarity to G. intermedius, Lactobacillus casei, Lactobacillus plantarum, and Acetobacter peroxydans.

Anti-microbial Activities Against Oral Microbes and Growth-Inhibitory Effect on Oral Tumor Cell of Extracts of Perilla and Mugwort (들깨잎과 쑥 추출물의 구강병 원인균에 대한 항균 및 KB 세포 증식 억제효과)

  • Jo, Min-Jung;Min, Kyung-Jin
    • Journal of Environmental Health Sciences
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    • v.33 no.2 s.95
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    • pp.115-122
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    • 2007
  • Methanol extracts of Perilla and Mugwort were stepwise extracted with hexane, chloroform, ethyl acetate, butanol and water. Anti-microbial activities and inhibitory effect on growth of oral tumor cell of each extract were investigated. Each extracts of Perilla and Mugwort were investigated to anti-microbial effects on oral microbes by means of agar diffusion method and MIC. These results suggest that the hexane extracts of Perilla and Mugwort have antimicrobial activities against S. mutans and potent inhibitory Effect to KB cell growth.