• Title/Summary/Keyword: agar dilution method

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Comparison of the E-Test with Agar Dilution Susceptibility Test by Using Bacteroides fragilis (Bacteroides fragilis의 E-test와 한천 평판 희석법에 의한 항균제 감수성상의 비교)

  • Kim, Hee-Sun;Kim, Sung-Kwang;Cha, Hwa-Sun
    • Journal of Yeungnam Medical Science
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    • v.10 no.1
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    • pp.135-143
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    • 1993
  • The susceptibilities of 45 clinical isolates of bacteroides frogilis to cefaclor, ciproflxacin and imipenem were determined by new method, E-test (AB Bidisk, Solna, Sweden) and were compared with those from conventional agar dilution method by using brain heart infusion, Mueller-Hinton and Wilkins Chalgren agar plates. And the susceptibility of 60 clinical isolates of Bacteroides fragilis group (B. fragilis 45 strains, B. distasonis 6 strains, B. ovatus 5 strains, B. thetaiotaomicron 4 strains) to 5 quinolones (ciprofloxacin, enoxacin, norfloxacin, ofloxacin, pefloxacin) were determined by in vitro agar dilution method. Compared with agar dilution MICs for B. fragilis 45 strains, 90.3% of E-test MICs were within ${\pm}$1 dilution of the agar dilutions, and 98.4% were within 2 dilutions. And there were little effect of different medium bases to determine MICs except Mueller-Hinton agar. On Mueller-Hinton agar, B. fragilis showed have or no growth activity. In vitro susceptibility of B. fragjlis group to quinolones, most of the test strains showed resistant patterns to quinolones except ofloxacin and there was little difference of susceptibility patterns between species of B. fragilis group.

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Antimicrobial susceptibility features of porcine respiratory bacterial pathogens by modified broth dilution method (돼지 호흡기질병 병인체의 항균제 감수성 조사)

  • 송동준;서동균;이춘식;배영찬;김원일;김봉환
    • Korean Journal of Veterinary Service
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    • v.23 no.1
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    • pp.19-28
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    • 2000
  • There are several main antibacterial susceptibility tests, such as agar dilution method, broth dilution method and disk diffusion technique. Especially, for minimal inhibitory concentration (MIC) test, agar dilution method has been widely used. But that method is so complicated and bothering that it's difficult to treat a large amount of strains. On the other hand, modified broth dilution method(add 1% glucose and 0.018% phenol red as a pH indicator to broth) is fast and easy to perform. Most of all, it can visualize the result by color. The MICs of 22 antibiotics Including penicillins, aminoglycosides, cephalothin, chloramphenicol, lincomycin, ceftiofur, vancomycin and quinolones, erythromycin, colistin. sul-fadimethoxine, trimethoprim for arcanobacterium pyogenes 14 strains, actinobacillus pleuropneu-moniae 41 strains and pasteurella multocida 37 strains, which were collected from porcine during 1996 ∼ 1999, were determined by modified broth dilution method. Actinobacillus pleuropneumoniae was highly susceptible to all kinds of quinolones such as ciprofloxacin, enrofloxacin and norfloxacin and to all aminoglycosides, like gentamicin, apramycin, kanamycin and ampicillin, cephalothin and ceftiofur. But It was quite resistant to solfadimethoxin, colistin and vancomycin. Pasteurella multocida was found to have high susceptibility to ampicillin, cephalothin, chlorampenicol and gentamicin but had mid-degree susceptibility to other aminoglycosides. In addition, it was susceptible to norfloxacin and nalidixic acid, but not to newer fluoroquinolone like ciprofloxacin and enrofloxacin and it was resistant to colistin and kanamycin. Arcanobacterium pyogenes was highly susceptible to most of quinolones such as cipoofloxacin, enrofloxacin and norfloxacin and gentamicin and penicillin G. But it also obtained high resistance against the early quinolone, nalidixic acid and aminoglycosides such as amikacin, apramycin and kanamycin and erythromycin, chlorampenicol, tetracyclin and vancomycin.

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A Study on the Antimicrobial Activity of Chitosan on the MRSA by Tube Dilution Technique and Agar Plate Smear Method (Tube Dilution Technique과 Ager Plate Smear Method에 의한 키토산의 MPSA 항미생물성)

  • Choi, Jeong-Im;Jeon, Dong-Won
    • Fashion & Textile Research Journal
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    • v.5 no.1
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    • pp.71-76
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    • 2003
  • Three different types of chitosan were prepared from red crab shells to study anti-microbial activity of chitosan on pathogenic bacteria, MRSA(Methicillin-resistant. Staphylococcus aureus), Water-insoluble chitosan, whose degree of deacetylation is kept over 90% and molecular weights are 20,000, 500,000, 150,000, 80,000, and 40,000, respectively. Water-soluble chitosan, whose degree of deacetylation is about 48% and molecular weights are 200,000 and 80,000. Water-soluble chitosan, whose degree of deacetylation is 82% and molecular weight is 3,900. The anti-microbial activities of three types of chitosan were investigated by Tube Dilution Technique(TDT) and Agar Plate Smear Method(APSM). And the following conclusions are made ; Chitosan having 5 different types of M.W chitosan (over 90% deacetylation) showed similar anti-microbial activities at over 0.05% concentration. Especially, chitosan having M.W 40,000 150,000 showed the excellent anti-microbial activity. The anti-microbial activity of chitosan was enhanced when the chitosan/acetic add solution was aged for 7days. The anti-microbial activity of chitosan was only shown at chitosan/acetic acid solution. The anti-microbial activity was not detected in chitosan solution dissolved in neutral pH water. Therefore, it can be concluded that the anti-microbial activity was due to NH3+ cationic ion of chitosan in acidic aqueous solution.

Isolation and Resistance Patterns of E. coli from Domestic Animals and Dranage in 1987 (가축 및 하천에서 분리한 대장균의 항균제내성)

  • Chung, Kyu-Sun;Lee, Yeon-Tae;Pio, Seog-Youn
    • YAKHAK HOEJI
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    • v.33 no.2
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    • pp.73-79
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    • 1989
  • One hundred and sixteen strains of E. coli isolated from drainage in Seoul city in 1987 and 104 strains of E. coli isolated from stools of domestic animals in suburb of Seoul in 1987 were examined for susceptibilities to 10 antimicrobial agents by the agar dilution method. The susceptibilities of the two groups to each antimicrobial agent were compared and correlations in the antimicrobial susceptibility of the 220 strains of E. coli among the 10 antimicrobial agents were also analyzed. The frequency of resistance strains was highest to tetracycline with 77%, and followed by chloramphenicol, streptomycin, ampicillin, kanamycin and cephalosporin in the descreasing order, tanging from 62% to 10%. Strains resistant to tobramycin, nalidixic acid, gentamicin and amikacin occupied 3 strains, 2 strains, 2 strains and 1 strain respectively.

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A new in vitro method for evaluating the antimicrobial activity of toothpaste

  • Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.46 no.2
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    • pp.94-97
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    • 2021
  • The purpose of this study was to introduce a new in vitro method for evaluating the antimicrobial activity of toothpaste, reflecting the actual toothbrushing time and the dilution of toothpaste by salivation. We designed three experimental groups and one negative control group. The experimental groups were (1) 90 μL of toothpaste + 10 μL 1X phosphate-buffered saline (PBS, 9/10 dilution group), (2) 50 μL of toothpaste + 40 μL 1X PBS (1/2 dilution group), and (3) 25 μL of toothpaste + 65 μL 1X PBS (1/4 dilution group). During toothbrushing, saliva is continuously secreted into the oral cavity and the toothpaste concentration is diluted over time during toothbrushing. Therefore, the 1/2 and 1/4 dilution experimental groups were added. The negative control group was toothpaste diluted 20,000-fold with 1X PBS. Miracle Fresh Doctor toothpaste and Streptococcus mitis KCOM 1350, Prevotella intermedia KCOM 1107, Fusobacterium nucleatum subsp. polymorphum KCOM 1322, and Aggregatibacter actinomycetemcomitans KCOM 1306 were used as the toothpaste and target bacterial strains, respectively. The number of bacterial cells plated on agar plates in the negative control group was 1,000 CFU. If the number of colonies on the experimental group plate was less than one, the treatment was considered to have > 99.9% bactericidal activity. These results suggest that this new in vitro method for antimicrobial evaluation could be used as the standard method for testing the antimicrobial activity of toothpaste.

Errors of Antibiotic Susceptibility Testing from Automated and Manual Systems in Clinical Isolates of Acinetobacter baumannii

  • Sung, Ji Youn;Oh, Ji-Eun;Kim, Eun Sun
    • Korean Journal of Clinical Laboratory Science
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    • v.45 no.1
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    • pp.21-25
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    • 2013
  • Acinetobacter baumannii is an aerobic, gram-negative and glucose-non-fermenting bacterium, which has emerged as a serious opportunistic pathogen. Many clinical microbiology laboratories use the Vitek 2 system for the routine antimicrobial susceptibility testing process, including testing on A. baumannii isolates. However, in case of amikacin, it is now recommended to perform additional antimicrobial susceptibility testing for A. baumannii strains due to the relatively lower minimum inhibitory concentration (MIC) in the Vitek 2 system compared to conventional reference methods. In our study, we assessed MIC for amikacin susceptibility testing of A. baumannii isolates in the Vitek 2 system, the agar dilution, Etest, and disk diffusion method. We collected 40 gentamicin-resistant, A. baumannii strains (amikacin MIC by Vitek 2:${\leq}2{\mu}g/mL$, 2 isolates; $4{\mu}g/mL$, 34 isolates; $8{\mu}g/mL$, 4 isolates) from a University hospital and compared the Vitek 2 system to other reference methods for testing susceptibility to amikacin. The Vitek 2 system showed major errors in all of the 40 isolates, yielding a low MIC. The results of our study strongly suggested that the Vitek 2 system was not a reliable method to test the MICs of gentamicin; ranging from ${\geq}16{\mu}g/mL$ for amikacin susceptibility. Other tests, such as agar dilution, Etest, or disk diffusion methods, should be paralleled to determine the MIC of amikacin in A. baumannii.

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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.

Bovine dermatophytosis and susceptibility of the isolates to antifungal drugs (소 피부사상균증의 발생상황 및 분리균의 약제감수성)

  • 오강희;박노찬;도재철;임소정;박진희
    • Korean Journal of Veterinary Service
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    • v.26 no.1
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    • pp.57-65
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    • 2003
  • The present study was conducted to examine the outbreak state of bovine dermatophytosis in 14 farms(4 dairy farms, 10 Korean indigenous cattle farms) in Gyeongbuk province from November 2000 to November 2001. The causative agents of dermatophytosis was identified by mycological examination. Antifungal susceptibility test of 26 isolates was performed by agar dilution method, using 5 antifungal drugs. Prevalence of bovine dermatophytosis was found to be 13.5%(90/665) in dairy cattle farms and 14.5%(220/1,520) in Korean indigenous cattle farms. The most common age at which this disease occurred was 2-12 months. This disease usually occurred from winter to spring and the occurrence subsequently decreased in the summer. But 4 Korean indigenous cattle farms with poorly hygienic status were occurred all the year round. The causative agent was identified as Trichophyton verrucosum exclusively in these case. Antifungal susceptibility test of T verrucosum (26 strains) was performed by agar dilution method, using 5 antifungal drugs including tolnaftate, griseofulvin, ketoconazole, amphotericin B and terbinafine. All isolates were highly sensitive to 5 antifungal drugs (geometric mean MICs 0.004∼0.032 $\mu\textrm{g}$/$m\ell$). The isolates were the most sensitive to especially tolnaftate.

Evaluation of Antimicrobial Activity of the Methanol Extracts from 8 Traditional Medicinal Plants

  • Kang, Chang-Geun;Hah, Dae-Sik;Kim, Chung-Hui;Kim, Young-Hwan;Kim, Eui-Kyung;Kim, Jong-Shu
    • Toxicological Research
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    • v.27 no.1
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    • pp.31-36
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    • 2011
  • The methanol extract of 12 medicinal plants were evaluated for its antibacterial activity against Gram-positive (5 strains) and Gram-negative bacteria (10 strains) by assay for minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC). The antibacterial activity was determined by an agar dilution method (according to the guidelines of Clinical and Laboratory Standard Institute). All the compounds (12 extracts) of the 8 medicinal plants (leaf or root) were active against both Gram-negative and Gram-positive bacteria. Gram-negative showed a more potent action than Gram positive bacteria. The MIC concentrations were various ranged from $0.6\;{\mu}g/ml$ to $5000\;{\mu}g/ml$. The lowest MIC ($0.6\;{\mu}g/ml$) and MBC ($1.22\;{\mu}g/ml$) values were obtained with extract on 4 and 3 of the 15 microorganisms tested, respectively.

Cytochrome b Gene-Based Assay for Monitoring the Resistance of Colletotrichum spp. to Pyraclostrobin

  • Dalha Abdulkadir, Isa;Heung Tae, Kim
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.616-628
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    • 2022
  • Resistance to pyraclostrobin due to a single nucleotide polymorphism at 143rd amino acid position on the cytochrome b gene has been a major source of concern in red pepper field infected by anthracnose in Korea. Therefore, this study investigated the response of 24 isolates of C. acutatum and C. gloeosporioides isolated from anthracnose infected red pepper fruits using agar dilution method and other molecular techniques such as cytochrome b gene sequencing, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and allele-specific polymerase chain reaction (PCR). The result showed that four isolates were resistant to pyraclostrobin on agar dilution method and possessed GCT (alanine) codon at 143rd amino acid position, whereas the sensitive isolates possessed GGT (glycine). Furthermore, this study illustrated the difference in the cytochrome b gene structure of C. acutatum and C. gloeosporioides. The use of cDNA in this study suggested that the primer Cacytb-P2 can amplify the cytochrome b gene of both C. acutatum and C. gloeosporioides despite the presence of various introns in the cytochrome b gene structure of C. gloeosporioides. The use of allele-specific PCR and PCR-RFLP provided clear difference between the resistant and sensitive isolates. The application of molecular technique in the evaluation of the resistance status of anthracnose pathogen in red pepper provided rapid, reliable, and accurate results that can be helpful in the early adoption of fungicide-resistant management strategies for the strobilurins in the field.