• Biomedical Science Letters
• /
• v.22 no.4
• /
• pp.167-173
• /
• 2016

The broth microdilution technique used to measure the minimal inhibitory concentration (MIC) of natural compounds against bacteria is problematic: it is difficult to visualize bacterial growth due to the color of the natural compound. Therefore, the use of a colorimetric method with a redox indicator by broth microdilution can simplify it and increase its objectivity. This study evaluated the usefulness of the colorimetric method in measuring the MIC of Phellinus baumii against methicillin-resistant Staphylococcus aureus (MRSA). The inhibition in disc diffusion method was observed from $8,192{\mu}g/mL$ P. baumii in all 10 MRSA isolates examined; however, the MIC ranges of the 10 MRSA isolates was $512{\sim}2,048{\mu}g/mL$ by broth microdilution using a colorimetric method; with the 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) indicator. In addition, the MIC of P. baumii by broth microdilution using MTT as indicator yielded excellent results. However, the 2, 3, 5-triphenyltetrazolium chloride (TTC) results could not be determined due to the color of the TTC indicator. The MICs of four antibiotics against MRSA using MTT or TTC were equal to those determined by visual interpretation. In conclusion, to evaluate the antibacterial effects of a natural compound, the broth microdilution technique is considered to be better than the disc diffusion method. Moreover, to resolve the problems caused by the colors of natural compounds, a colorimetric method such as that using MTT may be very valuable.

• International Journal of Oral Biology
• /
• v.36 no.4
• /
• pp.163-166
• /
• 2011

Oral viridans streptococci are recognized as one of the etiological agents of a variety of infectious diseases such as dental caries and infective endocarditis. Although antimicrobial susceptibility tests for these fastidious bacterial species are now established and standardized, a comparison between the broth microdilution and broth macrodilution tests has not previously been performed. This comparison was performed in the present study using the tests adopted by the Clinical and Laboratory Standards Institute (CLSI) and seven clinical isolates of oral viridans streptococcal strains. A modified broth macrodilution susceptibility test method was also included in this analysis, in which the media was not supplemented with horse blood. The susceptibility interpretation category agreements were measured at 83% (broth microdilution versus broth macrodilution) and 71% (broth microdilution versus modified broth macrodilution). The interpretation category agreement between the broth macrodilution and modified broth macrodilution tests was also 83%. These data indicate that the interpretation of antibiotic susceptibility test results for oral viridans streptococci are influenced by the methods used.

• Biomedical Science Letters
• /
• v.22 no.4
• /
• pp.174-183
• /
• 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.

• Mycobiology
• /
• v.37 no.3
• /
• pp.243-246
• /
• 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.

• Archives of Pharmacal Research
• /
• v.21 no.3
• /
• pp.278-285
• /
• 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.

• Mycobiology
• /
• v.37 no.1
• /
• pp.67-68
• /
• 2009

Antifungal activity of celery essential oil against Malassezia furfur was investigated using broth microdilution and vapor contact methods. Potent antifungal activity was evident using both methods. Fungicidal activity was revealed in the vapor contact method.

• Journal of Pharmacopuncture
• /
• v.19 no.1
• /
• pp.45-50
• /
• 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.

• Restorative Dentistry and Endodontics
• /
• v.19 no.2
• /
• pp.568-584
• /
• 1994

Bacteria have been regarded as major etiolgic factors in root canal infections. Infected root canal flora from thirteen patients who had visited to conservative department of Wonkwang dental hospital were cultured on blood agar plates. Cultued microorganisms were isolated and identified with Gram stain and biochemical tests using Vitek Systems(BioMeriux, MO, USA); Antibiotic susceptibillity was performed with disk diffusion and broth microdilution using Vitek Systems. Gram positive cocci(65 %) were predominant, which were composed of 6 Streptococcus viridans group, 5 Staph. spp., and 4 Enterococcus faecium, in the isolatd 23 strains. Gram negative rods (26 %) were the next common bacteria, which were composed of 5 non - fermentative Gram negative rods, and 1 Enterobacter cloacae. Most strains of S. viridans group and E. faecium were susceptible to antibiotics including penicillin. But strains of Staphylococcus spp. and non - fermentative Gram negative rods showed marked resistance to antibiotics except tetrancyclin and cefotaxime. Most results between disk diffusion and microdilution were all agreed, but the results of non - fermentative Gram negative rods were susceptible to cefotaxime in disk diffusion method but resistant in microdilution.

• Journal of Veterinary Clinics
• /
• v.22 no.4
• /
• pp.371-376
• /
• 2005

The aim of this study was to determine tile inhibitory effect of granitic powder against Microsporum canis. Fourteen strains of M. canis isolated from dgs and cats with fungal dermatitis and two strains isolated from humans were used in this study. The in vitro antifungal activities of granitic powder and 5 commercialized antifungal agents (terbinafine, itraconazole, ketoconazole, griseofulvin and fluconazole) were compared. The antifungal effect was measured by the broth microdilution method and was expressed as the minimal inhibitory concentration (MIC). The MIC value of the granitic powder was ranged from 31.3 to 250mg/ml. Terbinafine showed the lowest MIC value among the 5 commercial antifungal agents $(0.0078-0.125{\mu}g/ml)$, while fluconazole showed the highest MIC values $(125-1,000{\mu}g/ml)$. The MIC range of itraconazole, griseofuvin and ketoconazole were $0.125-0.5{\mu}g/ml\;0.625-5{\mu}g/ml$ and $10-40{\mu}g/ml$ respectively. The Geometric mean(GM) MIC values of terbinafine and ketoconazole against M. canis isolated from human were $0.0078{\mu}g/ml\;and\;10{\mu}g/ml$, respectively, while the GM MIC values of these agents against M. canis isolated from animals were $0.063{\mu}g/ml\;and\;31.4{\mu}g/ml$, respectively. Other antifungal agents did not show any significant differences in antifungal activity against M. canis of animal or human origin. Although granitic powder was shown to have antifungal activity, it was much lower than that of the 5 commercialized antifungal agents.

• Tuberculosis and Respiratory Diseases
• /
• v.64 no.6
• /
• pp.422-426
• /
• 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.