• Title/Summary/Keyword: Broth dilution method

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Effect of Sub-Minimal Inhibitory Concentration Antibiotics on Morphology of Periodontal Pathogens

  • Kwon, Ye Won;Lee, Si Young
    • International Journal of Oral Biology
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    • v.39 no.2
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    • pp.115-120
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    • 2014
  • Minimal inhibitory concentration (MIC) is the lowest concentration of antibiotics that inhibits the visible growth of a microorganism. It has been reported that sub-MIC of antibiotics may result in morphological alterations along with biochemical and physiological changes in bacteria. The purpose of this study was to examine morphological changes of periodontal pathogens after treatment with sub-MIC antibiotics. Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis were used in this study. The MIC for amoxicillin, doxycycline, metronidazole, penicillin, and tetracycline were determined by broth dilution method. The bacterial morphology was observed with bright field microscope after incubating with sub-MIC antibiotics. The length of A. actinomycetemcomitans and F. nucleatum were increased after incubation with metronidazole; penicillin and amoxicillin. P. gingivalis were increased after incubating with metronidazole and penicillin. However, F. nucleatum showed decreased length after incubation with doxycycline and tetracycline. In this study, we observed that sub-MIC antibiotics can affect the morphology of periodontal pathogens.

Synthesis of 1,3-Dioxolan-2-yliden Derivatives and Their Antifungal Activities (1,3-Dioxolan-2-yliden 유도체들의 합성과 항진균 활성)

  • 김영섭;김우정;김범태;박노균;박창식
    • YAKHAK HOEJI
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    • v.43 no.5
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    • pp.566-571
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    • 1999
  • (1H-1,2,4-Triazolyl) methy-4-(sub). phenyl-5-methyl-1,3-dioxolan-2-yliden (3) derivatives were synthesized and tested for their antifungal activities. The designed compounds with a 1,2,4-triazolylmethyl group at the 4-position of 1,3-dioxolan-2-yliden moiety were synthesized by reaction of difluorinated olefins(2) with (2R, 3R)-2-(2,4-dihalophenyl)-1-(1H-1,2,4-triazol-l-yl) butane-2,3-diol (1). These compounds were tested for in vitro antifungal activities against 16 fungi species. The MIC values were determined by the micro broth dilution method. In general, 1,3-dioxolan-2-yliden derivatives showed antifungal activities in vitro. Among them, (4R, 5R)-4-(2,4-difluorophenyl)-5-methyl-2-[1-(3,4-methylenedioxypheny)meth-ylidene)-1,3-dioxolon-4-yl(1H-1,2,4-triazollyl)methane showed superior antifungal activities to fluconazol and ketoconazol.

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Study on antifungal activity of herb oils against Trichophyton spp

  • Shin, Seung-Won;Kim, Ji-Hyun;Lim , Sook;Pyun, Mi-Sun
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.384.1-384.1
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    • 2002
  • The antifungal activities of the essential oils from Citrus borgamia. Ciderus atlantica. Cymbopogon ditratus, Eucalyptus globulus. Juniperus communis. Lavandula angustifolia. Melaeuca aterinfolia. Pelargonium graveolens. Pogestemon patchouli. Rosmarinus officinalis. Styrax tonkinensis. and Thymus vulgaris, which are recommended for the treatment of microbial infections in aromatherapy and complementary medicines. were tested against Trichophyton spp. The activities were measured by broth dilution method and disk diffusion assay. As the results, most of the test oils inhibited growth of T. tonsurans. T.mentagrophytes. T. ferugineum. and T. rubrum. Eapecially, the essential oils from C. atlantica. C. ditratus. e. globulus, and P. graveolens showed the strongest activity among the tested herb oils showing MICs between < 0.09 and 0.39 mg/ml.

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Antibacterial activity of Tonghyeonipal-dan against Methicillin-resistant Staphylococcus aureus (통현이팔단 에탄올 추출물의 Methicillin Resistant Staphylococcus aureus에 대한 항균활성)

  • KIM, In-Won;KANG, Ok-Hwa;KONG, Ryong;KWON, Dong-Yeul
    • The Korea Journal of Herbology
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    • v.30 no.5
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    • pp.15-21
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    • 2015
  • Objectives : Methicillin-resistantStaphylococcus aureus(MRSA) is a human pathogen. New antibacterial agents are needed to treat MRSA-related infections. This study investigated the antibacterial activity of EtOH 70% extracts ofTonghyeonipal-dan(THD) which prescription is composed of oriental medicine against MRSA.Methods : The antibacterial activity of THD was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method, Checkerboard dilution test, and Time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors were used.Results : The minimum inhibitory concentration (MIC) of THD is 1,000~2,000 μg/mL against MRSA. In the checkerboard dilution test, fractional inhibitory concentration index (FICI) of THD in combination with antibiotics indicated synergy or partial synergism againstS. aureus. Furthermore, a time-kill assay showed that the growth of the tasted bacteria was considerably inhibited after 24 h of treatment with the combination of THD with selected antibiotics. For measurement of cell membrane permeability, THD 500 μg/mL along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (TRIS) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide (NaN3) were used as an inhibitor of ATPase. TX, TRIS, DCCD and NaN3 cooperation againstS. aureusshowed synergistic action.Conclusions : Accordingly, antimicrobial activity of THD was affected by cell membrane and inhibitor of ATPase were assessed. These results suggest that THD has antibacterial activity, and that THD extract offers great potential as a natural antibiotic against MRSA.

Antibacterial Activity and Inhibition of Resistance in Methicillin-resistant Staphylococcus aureus by Maneung-hwan Ethanol Extract (만응환(萬應丸) 에탄올 추출물의 메티실린 내성 포도상구균에 대한 항균활성 및 내성억제 효과)

  • Na, Yong-su;Kim, Jong-gyu;Song, Yung-sun
    • Journal of Korean Medicine Rehabilitation
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    • v.30 no.1
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    • pp.31-45
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    • 2020
  • Objectives In this study, we investigated the antimicrobial activity of a 70% ethanol extract of Maneung-hwan (MEH), which is prescribed by practitioners of oriental medicine for use against methicillin-resistant Staphylococcus aureus (MRSA). Methods The antibacterial activity of MEH against MRSA strains was evaluated using the disc diffusion method, broth microdilution method (minimal inhibitory concentration, MIC), checkerboard dilution test, and time-kill test. The mechanism of action of MEH was investigated by bacteriolysis using detergents or ATPase inhibitors Additionally, mRNA and protein expression were investigated by quantitative reverse transcription-polymerase chain reaction and western blot assay, respectively. Results The MIC of MEH was 25~1,600 ㎍/mL against all the tested bacterial strains. We showed that MEH extract exerts strong antibacterial activity. In the checkerboard dilution test, the fractional inhibitory concentration index of MEH in combination with antibiotics indicated synergism or partial synergism against S. aureus. The time-kill study indicated that the growth of the tested bacteria was considerably inhibited after a 24-h treatment with MEH and selected antibiotics. To measure the cell membrane permeability, MEH (3.9 ㎍/mL) was combined with Triton X-100 (TX) at various concentrations N,N-dicyclohexylcarbodimide (DCCD) was also tested as an ATPase inhibitor. TX and DCCD cooperation against S. aureus exhibited synergistic action. Accordingly, the antimicrobial activity of MEH in the context of cell membrane rupture and ATPase inhibition was assessed. Additionally, the expression of genes and proteins associated with resistance was reduced after exposing MRSA to MEH. Conclusions These results suggest that MEH possesses antibacterial activity and acts as a potential natural antibiotic against MRSA.

A Study on Antibacterial Activity of Daehwanggeonwoo-san(Dahwangqianniu-san) Ethanol Extract against Methicillin-Resistant Staphylococcus Aureus (대황견우산(大黃牽牛散) 에탄올 추출물의 Methicillin 내성 Staphylococcus aureus에 대한 항균활성 연구)

  • Park, Ju-yeong;Na, Yong-su;Oh, Gong-cheon;Lee, Sang-mi;Choi, Byeong-kwon;Lee, Yoon-seung;Song, Yung-sun
    • Journal of Korean Medicine Rehabilitation
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    • v.28 no.2
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    • pp.21-35
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    • 2018
  • Objectives The objective of this study is to determine the antimicrobial effect of Daehwanggyeonu-san(Dahwangqianniu-san,DGE) and synergistic effects with antibiotics oxacillin, ampicillin, and gentamicin against Methicillin-Resistant Staphylococcus aureus(MRSA). Methods The antibacterial activity of DGE extract was evaluated againest MRSA strains by using the Disc diffusion method, broth microdilution method(minimal inhibitory concentration; MIC), checkerboard dilution test. The checkerboard dilution test was used to examined synergetic effect of oxacillin, ampicillin, gentamicin, ciprofloxacin with DGE extract. Results DGE showed antimicrobial activity against MRSA with an MIC value of $125{\sim}250{\mu}g/mL$. In the checkerboard test, the interation of DGE with all tested antibiotics produced almost synergy or partial synergy against MRSA. Conclusions This study shows that DGE reduced the MICs of several antibiotics tested, and a remarkable antibacterial effect of DGE, with membrane permeability enhancers and ATP synthase inhibitors. This study can be a valuable source for the development of a new drug with low MRSA resistance.

Enhanced Antimicrobial Activities and Physicochemical Characteristics of Isoliquiritigenin Encapsulated in Hydroxypropyl-β-Cyclodextrin (아이소리퀴리티게닌을 담지한 2-하이드록시프로필-베타사이클로덱스트린의 물리화학적 특성 및 항균활성 연구)

  • Kim, Hae Soo;Im, Na Ri;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.26 no.6
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    • pp.719-724
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    • 2015
  • Isoliquiritigenin (ILG) is a hydrophobic component in licorice and has a variety of pharmaceutical and biological activities. In this study, we prepared an isoliquiritigenin-hydroxypropyl-${\beta}$-cyclodextrin (ILG/HP-${\beta}$-CD) complex by freeze-drying method to enhance its water solubility. The complex was characterized by phase solubility studies, DSC, SEM, and 1H NMR. Antimicrobial activities against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) were evaluated by broth dilution method. The results showed that the stoichiometry of ILG/HP-${\beta}$-CD complex was 1 : 1. The antimicrobial activity of ILG/HP-${\beta}$-CD complex was higher than that of using free ILG against S. aureus and E. coli. Therefore, we suggest that ILG/HP-${\beta}$-CD complex may be used as a natural antiseptic and could potentially replace synthetic preservatives in cosmetic and food industries.

The Antimicrobial Activity of Medicinal Plants Extracts against Helicobacter Pylori (약용식물 추출물의 Helicobacter pylori에 대한 항균활성)

  • Lee, Jeong-Jun;Kim, Sung-Hoon;Chang, Byung-Sik;Lee, Joong-Bok;Huh, Chul-Sung;Kim, Tae-Jong;Baek, Young-Jin
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.764-770
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    • 1999
  • Antimicrobial activities were assayed through the hot-water extracts from 41 species of medicinal plants against Helicobacter pylori which is known as the ulcerogenic phathogen. Opuntia ficus-indica, Houttuynia cordata, Sinomenium acutum, and Coptis japonica showed the MIC at the concentrations less than 100 ppm, Pulsatilla koreana, Forsythia koreana, Rheum undulatum, and Perilla frutescens less than 200 ppm, Belamcanda chinensis, Arctium lappa, Cassia tora, Citrus tachibana, Siegesbeckia orientalis, and Caesalpinia sappan less than 300 ppm by the 2-fold dilution method. In disc method only three of them were confirmed to have antimicrobial activities which were increased in the order Perilla frutescens, Coptis japonica, Caesalpinia sappan. Three extracts were partitioned with chloroform, ethyl acetate and butanol in sequence and examined for the activity to inhibit H. pylori. The major ativities were observed in ethyl acetate fraction of Caesalpinia sappan, butanol fraction of Perilla frutescens, butanol and chloroform fraction of Coptis japonica. The partitioned fractions were found to have increased antimicrobial activities in all extracts. The experiments in which the extracts were added into urea R broth containing the crude urease derived from H. pylori resulted in the increase of pH and optical density at 560 nm to 8.15 and 1.7 respectively. Urease activity of H. pylori was inhibited over 80% by Caesalpinia sappan, Perilla frutescens, and Coptis japonica, of which Caesalpinia sappan suppressed up to 95%.

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Isolation and Numerical Identification of Streptomyces humidus strain S5-55 Antagonistic to Plant Pathogenic Fungi

  • Lim, Song-Won;Kim, Jeong-Dong;Kim, Biom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.16 no.4
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    • pp.189-199
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    • 2000
  • To search for the antifungal substances, various actino-mycete isolates were obtained from various soils of Korea using plate dilution method on the humic acid vitamin agar plates. In the screening procedures using a dual culture method, 32 actionomycete isolates were selected, which showed the inhibitory activity against mycelial growth of plant pathogenic fungi Altirnaria mali, Colletotrichum gloeosporides, Fusarium oxysporum f.sp. cucumerinum, Magnaporthe grisea, Phytophthora capsici, and Rhizoctonia solani. Bioassay of the crude extracts from culture filtrates and mycelial mets revealed that 12 antagonistic actionomycetes produced highly active antifungal substances. Actinomycete strain S5-55 which showed the substantial antifungal activity against the tested fungi was selected for production of the antifungal substances. Based on the cytochemical and morphological characteristics, strain S5-55 was identified as a Streptomyces species. The results of the numerical identification using the TAXON program confirmed that Streptomyces strain S5-55 was identical with Streptomyces humidus including in TAXON major cluster 19. The production of antifungal substance was most favorable when S. humidus strain S5-55 was cultivated for 10 dats on soluble starch broth supplemented with $K_2$HPO$_4$. The antifungal substances active against the plant pathogenic fungi P. capsici and M. grisea were partially purified using $\textrm{C}_{18}$ reversed-phase column chromatography.

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Drug Resistance and R-plasmid of E. coli Isolated from Patients, Domestic Animals and Drainages (환자, 가축 및 하천에서 분리한 대장균의 약제내성과 전달성 R-plasmid)

  • 김현주;정규선
    • YAKHAK HOEJI
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    • v.35 no.2
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    • pp.61-72
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    • 1991
  • These studies were made to assess the present stage of resistance to antibiotics, incidence and transferability of R-factors against E. coli. From March to July 1987, 59 strains of E. coli were isolated from specimens of patients collected at university hospitals in Seoul, 64 strains from stools of domestic animals and 66 strains from drainages in Seoul. These specimens were tested for resistance to 12 kinds of antimicrobial agents by means of agar dilution method. Using Muller-Hinton agar for the assay of drug resistance and tryptic soy broth as propagating medium for conjugation. The strains of E. coli were found to be resistant to one or more antibiotics and were considered to be potential donors of R-plasmid. The resistant strains of E. coli isolated from patients, domestic animals and drainages were found to be 55(93%), 33(52%) and 31(47%), respectively. Resistance to Tc, Ap and Cb was the highest in those isolated from patients and drainages, and resistance to Tc, Cm and Sm was the highest in those isolated from domestic animals. In the transfer test of drug resistance by conjugation method, 17 strains (47%) isolated from patients, 15(54%) isolated from domestic animals and 15(56%) isolated from drainages showed positive results, transperable resistant plasmid molecules with variable range in each strain.

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