• Title/Summary/Keyword: Microdilution assay

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Synthesis of Novel Halobenzyloxy and Alkoxy 1,2,4-Triazoles and Evaluation for Their Antifungal and Antibacterial Activities

  • Wan, Kun;Zhou, Cheng-He
    • Bulletin of the Korean Chemical Society
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    • v.31 no.7
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    • pp.2003-2010
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    • 2010
  • A new class of halobenzyloxy or alkoxy 1,2,4-triazoles and their hydrochlorides were synthesized through cyclization starting from commercially available phenylhydrazine. The structures were characterized by MS, IR and $^1H$ NMR spectra as well as elemental analyses. All the synthesized compounds were screened for their antibacterial activities in vitro against Staphylococcus aureus (ATCC29213), methicillin-resistant Staphylococcus aureus (N315), Bacillus subtilis, Escherichia coli (ATCC25922), Pseudomonas aeruginosa, Shigella dysenteriae, Eberthella typhosa, and antifungal activities against Candida albicans (ATCC76615), Aspergillus fumigatus by broth microdilution assay method. The results of preliminary bioassay indicated that 3-(2,4-difluorobenzyloxy)-1-phenyl-1H-1,2,4-triazole hydrochloride exhibited the best inhibitory activity with an MIC value of 56.25 ${\mu}M$ against P. aeruginosa superior to Chloramphenicol, and showed comparable activity with Chloramphenicol against E. coli (ATCC25922).

Activity of Essential Oil from Mentha piperita against Some Antibiotic-Resistant Streptococcus pneumoniae Strains and Its Combination Effects with Antibiotics

  • Choi, Sung-Hee;Shin, Seung-Won
    • Natural Product Sciences
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    • v.13 no.2
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    • pp.164-168
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    • 2007
  • To investigate natural antibiotics from plant essential oils and to evaluate their synergism with current antimicrobial drugs in inhibiting antibiotic-resistant strains of Streptococcus pneumoniae. The minimal inhibitory concentrations (MICs) of eleven plant essential oils and their main components were established for two antibiotic-susceptible and two antibiotic-resistant strains of S. pneumoniae, using broth microdilution tests. Potential synergism with oxacillin, norfloxacin, or erythromycin was evaluated using a checkerboard microtitre assay. Among the tested oils, Mentha piperita oil and its main component, menthol, exhibited the strongest inhibitory activities against all of the tested strains. The activity of antibiotics against antibiotic-resistant strains of S. pneumoniae was enhanced significantly by combination with Mentha piperita oils and its main component, menthol. In conclusion, the combination Mentha piperita essential oil or menthol with antibiotics could be used to reduce the effective dose of antibiotic and to modulate the resistance of S. pneumoniae strains.

The antibacterial effect of Pleurotus eryngii extracts on oral bacteria (새송이버섯 추출물이 구강세균에 작용하는 항균효과)

  • Chon, In-Young;Yu, Eun-Ji;Yu, Sang-Cheol;Lee, Ji-Youn;Jung, Sang-Hee;Oh, Tae-Jin
    • Journal of Korean society of Dental Hygiene
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    • v.18 no.1
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    • pp.9-18
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    • 2018
  • Objectives: Pleurotus eryngii is used both for edible and medicinal purposes, and has a physiological activity. The purpose of this study is to investigate the antibacterial effect of Pleurotus eryngii against six oral pathogens (Staphylococcus aureus, Streptococcus criceti, Streptococcus mutans, Streptococcus ratti, Streptococcus sobrinus, and Actinomyces viscosus). Methods: The antibacterial activities of various extracts of Pleurotus eryngii were examined by disc diffusion assay and minimum inhibitory concentration (MIC). The disc diffusion assay was performed by putting a paper disc soaked in extracts on plates inoculated bacterial cultures. The MIC of these extracts was determined by using a broth microdilution assay at a concentration ranging between 0.03 mg/ml to 15.00 mg/ml. The growth inhibition effect of extracts was measured at 600 nm for 24 hrs. Results: The antibacterial activity was confirmed against all six tested bacteria at Pleurotus eryngii ethyl acetate extract by the disc diffusion method. Acetone extract showed the antibacterial activity only against 4 strains containing Streptococcus criceti, Streptococcus mutans, Streptococcus ratti, and Actinomyces viscosus. In ethanol extract, no activity was observed against other strains except Staphylococcus aureus. MIC values of ethyl acetate extract were the same, 7.50 mg/ml in all tested bacteria. Conclusions: Pleurotus eryngii exhibited the antibacterial activity against oral pathogens (Staphylococcus aureus, Streptococcus criceti, Streptococcus mutans, Streptococcus ratti, Streptococcus sobrinus, and Actinomyces viscosus). Thus, Pleurotus eryngii may be considered as a natural antibacterial agent for treatment of dental diseases.

Evaluation of Antioxidant, Cytoprotective and Antimicrobial Properties of Polygoni multiflori Radix Extract, Fractions and Its Major Constituent (하수오 추출물, 분획물 및 주성분의 항산화, 세포 보호 및 항균 활성에 관한 평가)

  • Shin, Hyuk Soo;Kim, Minwoo;Song, Jerry;Lee, Junseok;Ha, Yoonjeong;Jeon, Young Hee;Kim, Ji Woong;Lee, Yun Ju;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.4
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    • pp.407-417
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    • 2018
  • In this study, the antioxidant, cytoprotective and antimicrobial activities of 50% ethanol extract of Polygoni multiflori Radix (PMR) and its ethyl acetate fraction were evaluated to confirm the applicability as a functional ingredient. The activities of the major constituent of PMR were verified and 2, 3, 5, 4′-tetrahydroxystilbene 2-O-${\beta}$-D-glucoside (THSG) was confirmed to be the main component of extract and fraction using HPLC-DAD, LC-EIS-MS analysis. The phenolic and THSG contents of the ethyl acetate fraction were 11.1- and 3.0-folds higher than those of the ethanol extract, respectively. As a result of the DPPH assay and that of luminol dependent chemiluminescence assay in $Fe^{3+}$-EDTA/H2O2 system. the ethylacetate fraction was superior to the ethanol extract in free radical and ROS scavenging activities. Especially, the ethyl acetate fraction and THSG exhibited the similar scavenging activity like L-ascorbic acid in ROS scavenging activity. The ethyl acetate fraction perceived the most potent cytoprotective effect against oxidative damage of erythrocytes induced by photosensitization reaction, followed by the ethanol fraction, THSG and that of (+)-${\alpha}$-tocopherol, which was used as a positive control. Antimicrobial activities were evaluated by disc diffusion and broth microdilution assay against S. aureus, E. coli, P. aeruginosa and C. albicans. In particular, the antibacterial activity of the extract and fraction against S. aureus was superior to that of methyl paraben. Taken together, our results suggest that PMR could be used as a natural ingredient for antioxidant, cytoprotective and antimicrobial activities.

Antimicrobial Effect of Coptidis rhizome Extract against Mutans Streptococci and Periodontopathogens

  • Park, Soon-Nang;Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.40 no.2
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    • pp.79-83
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    • 2015
  • The purpose of the study was to investigate the antimicrobial activity of the methanol extract of Coptidis rhizome against the type strains of cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus, and the periodontopathogens, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola and Aggregatibacter actinomycetemcomitans. The antimicrobial activities of the crude extract and the methanol extract fractions of Coptidis rhizome separated by silica gel chromatography were evaluated by determining the minimal bactericidal concentration (MBC) values, using the microdilution method. The cell viability test of the extracts of Coptidis rhizome on the KB cells was also studied by methyl thiazolyl tetrazolium (MTT) assay. Our results showed that the 11th fraction (F11) of the methanol extract had the greatest antimicrobial activity against the tested bacteria, with no associated cytotoxicity on the KB cells, upto a concentration of $50{\mu}g/ml$. These results suggest that the silica gel chromatography fraction F11 of the methanol extract of Coptidis rhizome, could be useful in the development of oral hygiene products as an antimicrobial agent for the prevention of dental caries and periodontal diseases.

Combined Effects of the Essential Oil from Pelargonium graveolens with Antibiotics against Streptococcus pneumoniae

  • Choi, Sung-Hee;Lim, Sook;Shin, Seung-Won
    • Natural Product Sciences
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    • v.13 no.4
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    • pp.342-346
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    • 2007
  • The antimicrobial activity of the essential oil from Pelargonium graveolens and its effects when it was combined with current antibiotics against antibiotic-resistant strains of Streptococcus pneumoniae were evaluated. The minimal inhibitory concentrations (MICs) of the essential oil fraction and the main components of this plant were determined for two antibiotic-susceptible and two antibiotic-resistant strains of S. pneumoniae using broth microdilution tests. The combined effects of the oil with erythromycin, norfloxacin, or oxacillin were evaluated using a checkerboard microtitre assay. The combination of the oil fraction of P. graveolens, or its main component, together with the antibiotics tested significantly lowered the MICs of the antibiotics against all of the tested strains with fraction inhibiting concentration indices (FICs) ranging from 0.16 to 1.50. In particular, the activity of norfloxacin against all of the tested strains of S. pneumoniae was enhanced significantly by combination with citronellol. In conclusion, the combination of P. graveolens oil with antibiotics could be used to reduce the effective dose of antibiotic and to modulate the resistance of S. pneumoniae strains.

Opposite Effects of Vitamin C and Vitamin E on the Antifungal Activity of Honokiol

  • Sun, Lingmei;Ye, Xiaolong;Ding, Dafa;Kai, Liao
    • Journal of Microbiology and Biotechnology
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    • v.29 no.4
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    • pp.538-547
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    • 2019
  • The aim of the present study was to evaluate the effects of two well-known natural antioxidants, vitamin C (VC) and vitamin E (VE), on the antifungal activity of honokiol against Candida albicans. The broth microdilution method was employed to test the antifungal activities of honokiol with or without antioxidants in the medium against C. albicans strain. Intracellular reactive oxygen species and lipid peroxidation were determined by fluorescence staining assay. Mitochondrial dysfunction was assessed by detecting the mitochondrial DNA and the mitochondrial membrane potential. We observed that VC could significantly potentiate the antifungal activities of honokiol while VE reduced the effectiveness of honokiol against C. albicans. In addition, VC accelerated honokiol-induced mitochondrial dysfunction and inhibited glycolysis leading to a decrease in cellular ATP. However, VE could protect against mitochondrial membrane lipid peroxidation and rescue mitochondrial function after honokiol treatment. Our research provides new insight into the understanding of the action mechanism of honokiol and VC combination against C. albicans.

Synergy effect of legal highs with antibiotics (Legal High Plants와 항생제의 항균활성 비교)

  • Jung, Son-Hyo;You, Seon-Hee;Park, Cho-Hi
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.6
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    • pp.1635-1645
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    • 2020
  • In this study would like to find extending or increasing the efficacy of the antibiotic substance for the strains with resistance to antibiotics or persister cells by inhibition of the resistance. This study was used different species of 'legal high' plants leaves from Leonotis leonurus, Mitragyna speciosa, and seeds from Ipomoea murucoides with antibiotics which are Amoxicillin, Chloramphenicol, Ciprofloxacin, Kanamycin, Oxacillin, and Vancomycin. Legal highs were extracted with methanol. Minimum inhibitory concentration(MIC) testing for a range of antibiotics with extracts of plant was fulfilled by broth dilution methods. In this essay, it was determined in a microdilution assay utilizing suspended in ISB up to a final concentration of 512㎍/ml in 96 wells microtitre plates, threefold and serial dilutions. After that, the microplates were kept in incubator between 35℃ and 37℃ for overnight. Leonotis leonurus, Mitragyna speciosa, and Ipomoea murucoides of Legal highs (512㎍/ml) investigated small activity to inhibit against pathogens which are susceptible Staphylococcus aureus, resistant Staphylococcus aureus, susceptible Enterococcus faecalis, resistant Escherichia coli.

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.

Chemical Composition and Antimicrobial Efficacy of Helminthostachys zeylanica against Foodborne Bacillus cereus

  • Yenn, Tong Woei;Ring, Leong Chean;Zahan, Khairul Azly;Rahman, Muhammad Sharir Abdul;Tan, Wen-Nee;Alaudin, Bintul Jauza' Shaik
    • Natural Product Sciences
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    • v.24 no.1
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    • pp.66-70
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    • 2018
  • Helminthostachys zeylanica is a rare plant grows in lightly shaded areas. The fern was traditionally used as antipyretic and antiphlogistic agents. This study was aimed to evaluate the antibacterial potential of H. zeylanica on foodborne Bacillus cereus. The chemical composition of its ethanolic extract was also determined. The plant samples were collected at Kampung Kebun Relong, Kedah, Malaysia. The ethanolic extract showed significant inhibitory activity on B. cereus with a sizeable clear zone detected on disc diffusion assay. On broth microdilution assay, the MIC of the extract on B. cereus was 6.25 mg/ml and the MBC was 12.5 mg/ml. The inhibitory activity of the extract on B. cereus was bactericidal. In the growth dynamic study, the antibacterial efficacy of the extract was concentration dependent, where a lower colony forming unit count was obtained with increased extract concentration. The SEM micrograph of extract treated B. cereus cells showed invaginations of cell wall. The bacterial cell structure collapsed after 24 h exposure to the extract. The GCMS analysis of the extract showed that the major constituents of the extract were phenol (36.26%) and quercetin (29.70%). This study is important as it shows the potential use of H. zeylanica as an effective agent to control B. cereus related infections.