• Title/Summary/Keyword: Microdilution assay

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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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Evaluation and Optimization of a Serum-based Minimum Inhibitory Concentration Assay to Caspofungin in Candida albicans Clinical Isolates

  • Yoo, Young Bin;Kim, Sung-Soon;Kim, Young Kwon;Kim, Sunghyun
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.174-183
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    • 2016
  • In the present study, a serum-based minimum inhibitory concentration (MIC) testing to caspofungin was optimized and evaluated to solve the limitations of the conventional Clinical and Laboratory Standards Institute (CLSI) guideline-based antifungal agent MIC test and the usefulness of this testing for clinical application was determined. A total of 105 Candida albicans clinical isolates were used for measuring MIC to caspofungin. Results showed that growth characteristics were different according to types of serum and the mouse serum was the most suitable for this assay. In order to measure the optimal concentration of mouse serum, 0 to 100% mouse serum were added to the media during fungal culture. The optimal concentration of serum was 50% when consideration of antifungal agent administration and inoculum size, serum components and ease of hyphae separated, and the consideration of the degree of growth. In comparison of the usefulness between the conventional Alamar-modified broth microdilution MIC assay and 50% mouse serum-based MIC testing, the range of $MIC_{80}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.42{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $2.0{\sim}32.0{\mu}g/mL$ (SD ${\pm}9.01{\mu}g/mL$). The range of $MIC_{50}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.40{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $1.0{\sim}16.0{\mu}g/mL$ (SD ${\pm}2.36{\mu}g/mL$). The MICs of 50% mouse serum-based MIC testing was increased by up to 4 to 64 times than Alamar-modified broth microdilution MIC assay. In conclusion, a 50% mouse serum-based MIC assay was more useful for measuring MIC in Candida albicans clinical isolates than conventional colorimetric broth microdilution MIC testing.

Antifungal Activity of Bee Venom and Sweet Bee Venom against Clinically Isolated Candida albicans

  • Lee, Seung-Bae
    • Journal of Pharmacopuncture
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    • v.19 no.1
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    • pp.45-50
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    • 2016
  • Objectives: The purpose of this study was to investigate the antifungal effect of bee venom (BV) and sweet bee venom (SBV) against Candida albicans (C. albicans) clinical isolates. Methods: In this study, BV and SBV were examined for antifungal activities against the Korean Collection for Type Cultures (KCTC) strain and 10 clinical isolates of C. albicans. The disk diffusion method was used to measure the antifungal activity and minimum inhibitory concentration (MIC) assays were performed by using a broth microdilution method. Also, a killing curve assay was conducted to investigate the kinetics of the anti-fungal action. Results: BV and SBV showed antifungal activity against 10 clinical isolates of C. albicans that were cultured from blood and the vagina by using disk diffusion method. The MIC values obtained for clinical isolates by using the broth microdilution method varied from $62.5{\mu}g/mL$ to $125{\mu}g/mL$ for BV and from $15.63{\mu}g/mL$ to $62.5{\mu}g/mL$ for SBV. In the killing-curve assay, SBV behaved as amphotericin B, which was used as positive control, did. The antifungal efficacy of SBV was much higher than that of BV. Conclusion: BV and SBV showed antifungal activity against C. albicans clinical strains that were isolated from blood and the vagina. Especially, SBV might be a candidate for a new antifungal agent against C. albicans clinical isolates.

In vitro Antifungal Activity of Limonene against Trichophyton rubrum

  • Chee, Hee-Youn;Kim, Hoon;Lee, Min-Hee
    • Mycobiology
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    • v.37 no.3
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    • pp.243-246
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    • 2009
  • In this study, the antifungal activities of limonene against Trichophyton rubrum were evaluated via broth microdilution and vapor contact assays. In both assays, limonene was shown to exert a potent antifungal effect against T. rubrum. The volatile vapor of limonene at concentrations above $1{\mu}l$/800 ml air space strongly inhibited the growth of T. rubrum. The MIC value was 0.5% v/v in the broth microdilution assay. The antifungal activity of limonene against T. rubrum was characterized as a fungicidal effect.

Detection of Clarithromycin-resistant Strains from Clinical Isolates of Mycobacterium abscessus (Clarithromycin 약제에 대한 Mycobacterium abscessus 임상 내성 균주 분리)

  • Lee, Seung Heon;Park, Young Kil;Ryo, Sung Weon;Shim, Myung Sup;Lew, Woo Jin;Kim, Hee Jin
    • Tuberculosis and Respiratory Diseases
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    • v.64 no.6
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    • pp.422-426
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    • 2008
  • Background: Mycobacterium abscessus is the most pathogenic and drug-resistant rapid-growing mycobacterium. Clarithromycin or azithromycin are the only regular oral antimycobacterial agents that have an effect on M. abscessus. We tried to detect the clarithromycin-resistant strains from the clinical isolates of M. abscessus. Methods: We tried to isolate the clarithromycin-resistant strains from 220 clinical isolates of M. abscessus by performing using reverse hybridization assay (RHA) and the broth microdilution test (BMT). Results: Seven resistant strains (3.2%) from all the tested clinical isolates were detected by BMT. Three of these resistant strains were also detected by RHA and it was confirmed that they had point mutants. Conclusion: These results showed that clarithromycin resistance in M. abscessus clinical isolates is related to a point mutation and other unknown mechanisms.

Assays of Residual Antibiotics after Treatment of γ-ray and UV Irradiation (감마선과 UV 조사에 의한 항생제 분해)

  • Shin, Ji-Hye;Nam, Ji-Hyun;Yu, Seungho;Lee, Myunjoo;Lee, Dong-Hun
    • Journal of Radiation Industry
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    • v.4 no.1
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    • pp.39-45
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    • 2010
  • The pollution of antibiotics is a major cause of spreading antibiotics resistant bacteria in the environment. Applications of ozonation, UV, and ${\gamma}-ray$ irradiations have been introduced to remove antibiotics in the effluents from wastewater treatment system. In this study, we compared the chemical (HPLC) and biological (antimicrobial susceptibility test, AMS) assays in measuring of the concentrations of residual antibiotics after ${\gamma}-ray$ and UV irradiation. Most samples were degraded by ${\gamma}-ray$ irradiation (1~2 kGy). However, lincomycin and tetracycline were not degraded by UV irradiation. The concentration of residual antibiotics, that was treated with ${\gamma}-ray$ and UV irradiation, measuring by bioassay was similar to HPLC. The concentrations of ${\gamma}-ray$ irradiated cephradine measured by AMS test were 2 times higher than that of HPLC assay, indicating AMS test is more sensitive than HPLC assay. These results indicate that ${\gamma}-ray$ irradiation technique is more useful than UV irradiation, and biological assay is more useful to detect the antibiotics and toxic intermediates in antibiotics degradation.

Endophytic Diaporthe sp. ED2 Produces a Novel Anti-Candidal Ketone Derivative

  • Yenn, Tong Woei;Ring, Leong Chean;Nee, Tan Wen;Khairuddean, Melati;Zakaria, Latiffah;Ibrahim, Darah
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1065-1070
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    • 2017
  • This study aimed to examine the anti-candidal efficacy of a novel ketone derivative isolated from Diaporthe sp. ED2, an endophytic fungus residing in medicinal herb Orthosiphon stamieus Benth. The ethyl acetate extract of the fungal culture was separated by open column and reverse phase high-performance liquid chromatography (HPLC). The eluent at retention time 5.64 min in the HPLC system was the only compound that exhibited anti-candidal activity on Kirby-Bauer assay. The structure of the compound was also elucidated by nuclear magnetic resonance and spectroscopy techniques. The purified anti-candidal compound was obtained as a colorless solid and characterized as 3-hydroxy-5-methoxyhex-5-ene-2,4-dione. On broth microdilution assay, the compound also exhibited fungicidal activity on a clinical strain of Candida albicans at a minimal inhibitory concentration of $3.1{\mu}g/ml$. The killing kinetic analysis also revealed that the compound was fungicidal against C. albicans in a concentration- and time-dependent manner. The compound was heat-stable up to $70^{\circ}C$, but its anti-candidal activity was affected at pH 2.

Antibacterial Activity of Ethanol Extract and Fraction of Sasa borealis (조릿대 Ethanol 추출물 및 분획물의 항균 효과)

  • Jang, Mi-Ran;Lee, Da-Uhm;Kim, Gun-Hee
    • Korean journal of food and cookery science
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    • v.26 no.6
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    • pp.848-852
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    • 2010
  • This study investigated the antibacterial activities of extracts and fractions of Sasa borealis against eight bacteria (Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Psedomonas aerginosa, Salmonella choleraesuis, Serratia marcescens and Vibrio vulnificus) by broth dilution assay. Using survival curves, the kinetics of bacterial inactivation upon exposure to the extracts and fractions were followed for 24 h. In this same manner, MIC (minimum inhibitory concentration) values were determined by broth microdilution assay and then confirmed to be the extract concentrations that inhibited bacterial growth. Sasa borealis extracts showed antibacterial activities against all tested bacteria. In particular, all tested fractions of Sasa borealis had stronger activities than 70% ethanol extract. MIC of Sasa borealis extract was determined to be 5 mg/mL against Salmonella choleraesuis. All fractions of Sasa borealis extract had extremely strong antibacterial activities. MIC of fractions were determined to be 0.03~2.5 mg/mL. These results suggest that the extracts and fractions of Sasa borealis effectively inhibited bacterial growth and thus are useful as natural antibacterial agents.