• Title/Summary/Keyword: Broth microdilution method

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Evaluation of the Antibacterial Effects of Phellinus baumii Extract on Methicillin-Resistant Staphylococcus aureus by Using Broth Microdilution Based on a Colorimetric Method

  • Shin, Kyeong Seob;Jo, Mi Young;Hong, Seung Bok
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.167-173
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    • 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.

Comparison of Three Antibiotic Susceptibility Tests for Viridans Group Streptococci

  • Kim, Yeon-Hee;Lee, Si-Young
    • International Journal of Oral Biology
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    • v.36 no.4
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    • pp.163-166
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    • 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.

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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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.

INDENTIFICATION AND ANTIBIOTIC SUSCEPTIBILITY TEST OF MICROORGANISMS ISOLATED FROM INFECTED ROOT CANALS (감염근관에서 분리한 세균의 동정 및 항생제 감수성 검사)

  • Ji, Jung-Ho;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.19 no.2
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    • pp.568-584
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    • 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.

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Antifungal Activity of 5 Antifungal Agents and Granitic Powder on Microsporum canis (Microsporum canis에 대한 5가지 항진균제와 맥반석 가루의 항진균 효과)

  • Kang Tae-hyung;Lee Jeong-chi;Won Young-ho;Oh Seok-il;Lee Chung-gil;Lee Chai-yong
    • Journal of Veterinary Clinics
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    • v.22 no.4
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    • pp.371-376
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    • 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.

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).

Antimicrobial Susceptibility of Bordetella bronchiseptica Isolates from Pigs (돼지에서 분리한 Bordetella bronchiseptica의 항생제 감수성)

  • 조정곤
    • Journal of Veterinary Clinics
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    • v.15 no.1
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    • pp.30-35
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    • 1998
  • Susceptibility or resistance of the Bordetella bronchiseptica was determined by using the broth microdilution method. Each Bordetella bronchiseptica was tested fur sutraceptluility with 7 different antimicrobial agents.4 high prevalence of resistance (greater than 80 %) was found when Bordetella bronchiseptica was tested with amikacin (AK), ciprofloxacin ILrJl gentamlcin IGAgf, kanamycin (Kbf) and tobramycin (TM). However, the Bordetella bronchiseptica were sensitive to chloramphenicol (CP) and tetracycline (TC). Three different combination of drug resistance were observed : AM-CF-GM-TM (1 strain), AM-CF-GM- KM-TM (45 strains), and AM-CF-GM-KM-TM-CP (3 strains). The MICs against Bordetella bronchiseptica were 0.13 to >2.00 for TC, 4 to >648 for CP, 8 to >128 for CF 16 to >256 for GM, 16 to >256 for TM, 32 to >256 for KM and 64 to 256 for AM.

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Antimicrobial Resistance of Seventy Lactic Acid Bacteria Isolated from Commercial Probiotics in Korea

  • Eunju Shin;Jennifer Jaemin Paek;Yeonhee Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.4
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    • pp.500-510
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    • 2023
  • In this study, lactic acid bacteria were isolated from 21 top-selling probiotic products on Korean market and their antimicrobial resistance were analyzed. A total 152 strains were claimed to be contained in these products and 70 isolates belonging to three genera (Bifidobacterium, Lactobacillus, and Lactococcus) were obtained from these products. RAPD-PCR showed diversity among isolates of the same species except for two isolates of Lacticaibacillus rhamnosus from two different products. The agar dilution method and the broth dilution method produced different MICs for several antimicrobials. With the agar dilution method, five isolates (three isolates of Bifidobacterium animalis subsp. lactis, one isolate of B. breve, one isolate of B. longum) were susceptible to all nine antimicrobials and 15 isolates were multi-drug resistant. With the broth microdilution method, only two isolates (one isolate of B. breve and one isolate of B. longum) were susceptible while 16 isolates were multi-drug resistant. In this study, only two AMR genes were detected: 1) lnu(A) in one isolate of clindamycin-susceptible and lincomycin-resistant Limosilactobacillus reuteri; and 2) tet(W) in one tetracycline-susceptible isolate of B. longum B1-1 and two tetracycline-susceptible isolates and three tetracycline resistant isolates of B. animalis subsp. lactis. Transfer of these two genes via conjugation with a filter mating technique was not observed. These results suggest a need to monitor antimicrobial resistance in newly registered probiotics as well as probiotics with a long history of use.