• Title/Summary/Keyword: agar dilution test

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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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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.

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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Comparative Study Using MIC Test for Cefazedone and other First Generation Cephalosporins (1세대 세파계 항생제인 Cefazedone의 MIC Test를 통한 비교 연구)

  • Bok, Hae-Sook;Kim, Myung-Min;Choi, Kyung-Eob
    • Korean Journal of Clinical Pharmacy
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    • v.5 no.2
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    • pp.13-16
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    • 1995
  • The Minimum Inhibitory Concentration (MIC) of a first-generation cephalosporin derivative, Cefazedone (CZD; $PAZERON^R$ inj.) was determined by the two-fold serial agar dilution method. The in-vitro antibacterial activity of CZD against a wide variety of clinical isolates was compared with those of other first generation cephalosporins such as Methylol Cephalexin (CEX), Cefazolin (CEZ), Cefadroxil (CDX), Cephradine (CED), Ceftezol (CTZ) and one of second generation cephalsporin antibiotics, Cefotaxime (CTX). CZD had the most potent inhibitory effect against Gram-positive strains, when compared to the first-generation cephalosporin antibiotics tested in this study and CTX. The geometric MIC mean of CZD for Gram-positive strains was calculated as 0.386 kg/m{\ell}$, and those of CEX, CEZ, CDX, CTZ, CED, and CTX were 6.073, 0.894, 3.399, 0.748, 7.884 and 1.502 $kg/m{\ell}$, respectively. In addition, the geometric mean of CZD for staphylococclJs aureus strains was obtained as 0.340 $kg/m{\ell}$ and those of CEX, CEZ, CDX, CTZ, CED, and CTX 6.145, 0.534, 4.126, 0.442, 10.51, and 2.500 $kg/m{\ell}$, respectively. Against Gram-negative strains, CZD showed better antibacterial activity than CEZ, CDX, CTZ, and CED.

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Drug susceptibility of bacteria and M pachydermatis isolated from canine external ear canals (개 외이도에서 분리한 세균 및 M pachydermatis의 약제감수성에 대하여)

  • Kim, Ki-hyang;Choi, Won-phil;Yeo, Sang-geon
    • Korean Journal of Veterinary Research
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    • v.39 no.3
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    • pp.560-565
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    • 1999
  • The present work was conducted to investigate the drug susceptibility of microorganisms isolated from canine external ear canals. Antifungal susceptibility test of M pachydermatis (17 strains) was perfomed by agar dilution method, using 11 antifungal drugs including amphotericin B(A), nystatin(N), pimaricin(P), griseofulvin(G), bifonazole(B), clotrimazole(C), miconazole(M), econazole(E), ketoconazole(K), tolnaftate(T), 5-fluorocytosine(F). All isolates were highly sensitive to K, M, T(geometric mean MIC ; GM $MIC{\leq}0.16{\mu}g/ml$) but they weren't sensitive to P, F and G(GM $MIC{\geq}92.37{\mu}g/ml{\sim}{\geq}128{\mu}g/ml$). Antibacterial susceptibility test against 119 isolates of bacteria was performed by agar dilution method, using 9 antibacterial drugs including erythromycin(ET), chloramphenicol(CP), gentamycin(G), vancomycin(V), ampicillin(AP), amoxacillin(AX), chlortetracycline(CT), ciprofloxacin(CF), enrofloxacin(EF). All isolates of Staphylococcus spp(101 strains) were highly sensitive to EF, CF, G(GM MIC $0.33{\sim}1.47{\mu}g/ml$). In other gram positive cocci(4 strains), they were highly sensitive to EF, CF, V(GM MIC $1{\sim}4.76{\mu}g/ml$) and CT(GM MIC 1 UFL unit/ml). In gram positive rods(13 strains), they were highly sensitive to EF, CF, G(GM $MIC{\leq}0.19{\sim}1{\mu}g/ml$). In Pseudomonas aeruginosa(1 strain), it was highly sensitive to AX, EF, ET, CF(GM MIC $0.06{\sim}1{\mu}g/ml$) and CT(GM MIC 1 UFL unit/ml). All isolates weren't sensitive to AP(GM MIC 16~>$32{\mu}g/ml$).

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Antimutagenic and Antibacterial Activities of Korean and American Propolis (한국산과 미국산 프로폴리스의 항돌연변이 및 항균효과)

  • Jang, Il-Woong;Park, Jeong-Seob;Kwon, Hyoung-Cheol;Jung, Mun-Yhung;Choi, Dong-Seong
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.694-699
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    • 2009
  • The antimutagenic activities of ethanol extracts of Korean and American propolis were tested using Salmonella Typhimurium TA98 with two indirect mutagens of 3-amino-1,4-dimethyl-5H-pyrido [4,3-b]indole (Trp-P-1) and 2-aminoanthracene (2-AA) with S9 mix. Additionally, their antimicrobial activities against acne-related pathogenic strains of Propionibacterium acnes, Staphylococcus Epidermidis, Staphylococcus aureus and Pseudomonas aeruginosa were evaluated using both paper disk method and agar dilution method. Ethanol extracts of Korean and American propolis showed strong inhibitory effects, in a dose dependant manner, against the mutagenicities induced by Trp-P-1 and 2-AA. The antimutagenic effect of ethanol extracts of Korean propolis showed significantly higher protective activity than that of American propolis against the Trp-P-1 induced mutagenicity of S. Typhimurium TA98 at the lower concentration ($1-10\;{\mu}g$), but significantly lower protective activity at the higher concentration ($50-200\;{\mu}g$). The antimutagenic effect of ethanol extract of Korean propolis showed significantly higher protective activity than that of American propolis against the 2-AA induced mutagenicity at the concentration of $1\;{\mu}g$, but significantly lower protective activity than that of the American at the higher concentration ($50-200\;{\mu}g$). Both extracts showed strong antimicrobial activities against all the acne-related pathogens tested, with minimal inhibitory concentration (MIC) values in the range $1,500-5,000\;{\mu}g/mL$.

Fungicide selections for control of chili pepper stem rot caused by Sclerotium rolfsii using an agar dilution method

  • Lee, Soo Min;Min, Jiyoung;Kim, Heung Tae
    • Korean Journal of Agricultural Science
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    • v.44 no.3
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    • pp.339-347
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    • 2017
  • Sclerotium rolfsii causing southern blight on numerous vegetable and fruit crops was isolated from stems of chili peppers showing wilting symptoms. The pathogen was identified by morphological observation and DNA sequencing analysis of ITS region. To select an effective fungicide for control of southern blight, we investigated the inhibition efficacy of thirty fungicides included in nine groups of fungicides with different mechanisms of action. A fungal growth inhibition assay was conducted through an agar dilution method by using mycelial discs and sclerotia of the pathogen as inoculum, respectively. When mycelial discs were used as an inoculum, several fungicides showed good inhibitory activity against the mycelial growth of S. rolfsii 12-6. All DMI fungicides tested had a good inhibition except for prochloraz which had low inhibitory effect. All strobilurin fungicides tested except for kresoxim-methyl and all SDHI fungicides tested except for boscalid and fluopyram, had a good inhibition. Also, fludioxonil, a protective fungicide and fluazinam had a good inhibitory effect. Interestingly, when sclerotia were used as an inoculum, inhibition efficacy was increased for fluopyram, a SDHI fungicide, and for some protective fungicides such as propineb, chlorothalonil, dithianon, and folpet. All the fungicides selected in this study should be tested in the field for their control activities against stem rot for practical use in chili pepper cultivation.

Riboflavin Inhibits Growth of Helicobacter pylori by Down-regulation of polA and dnaB Genes

  • Kwon, Hye Jin;Lee, Min Ho;Kim, Hyun Woo;Yang, Ji Yeong;Woo, Hyun Jun;Park, Min;Moon, Cheol;Kim, Sa-Hyun;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.26 no.4
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    • pp.288-295
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    • 2020
  • Infection of Helicobacter pylori on gastric mucosa is associated with various gastric diseases. According to the WHO, H. pylori causes gastric cancer and has been classified as a class I carcinogen. Riboflavin is an essential vitamin which presents in a wide variety of foods. Previous studies have shown that riboflavin/UVA was effective against the growth inhibition of Staphylococcus aureus, S. epidermidis and multidrug-resistant Pseudomonas aeruginosa and had the potential for antimicrobial properties. Thus, we hypothesized that riboflavin has a potential role in the growth inhibition of H. pylori. To demonstrate inhibitory concentration of riboflavin against H. pylori, we performed agar and broth dilution methods. As a result, we found that riboflavin inhibited the growth of H. pylori. The MIC was 1 mM in agar and broth dilution test. Furthermore, to explain the inhibitory mechanism, we investigated whether riboflavin has an influence on the replication-associated molecules of the bacteria using RT-PCR to detect mRNA expression level in H. pylori. Riboflavin treatment of H. pylori led to down-regulation of polA and dnaB mRNA expression levels in a dose dependent manner. After then, we also confirmed whether riboflavin has cytotoxicity to human cells. We used AGS, a gastric cancer cell line, and treated with riboflavin did not show statistically significant decrease of cell viability. Thus, these results indicate that riboflavin can suppress the replication machinery of H. pylori. Taken together, these findings demonstrate that riboflavin inhibits growth of H. pylori by inhibiting replication of the bacteria.

Standardization of an enzyme-linked immunosorbent assay for detection of antibody to avian reticuloendotheliosis virus (세망내피증 바이러스 항체검출을 위한 ELISA 표준화)

  • Sung, Haan Woo;Lee, Su Jeong
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.569-574
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    • 2005
  • Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.