scholarly journals In Vitro Antifungal Susceptibility Of Griseofulvin, Fluconazole, Itraconazole And Terbinafine Against Clinical Isolates Of Trichophyton Rubrum And Trichophyton Mentagrophytes

2014 ◽  
Vol 2 (1) ◽  
Author(s):  
K R Reddy ◽  
S Ram Reddy
Keyword(s):  
Trichophyton Rubrum ◽  
Clinical Laboratory ◽  
Antifungal Susceptibility ◽  
Trichophyton Mentagrophytes ◽  
Drug Susceptibility ◽  
Antifungal Drugs ◽  
Clinical Isolates ◽  
National Committee ◽  
In Vitro Antifungal Susceptibility

Investigations on antifungal drug susceptibility were carried out on 90 clinical isolates of Trichophyton rubrum, and Trichophyton mentagrophytes with four antifungal drugs, namely griseofulvin, fluconazole, itraconazole and terbinafine as suggested by National Committee for Clinical Laboratory Standards (NCCLS) M27–A (1997) document by broth macrodilution method to standardize in vitro antifungal susceptibility testing and to find out the Minimum Inhibitory Concentration (MIC) of the drugs. In this study, terbinafine was found to be the most efficient drug for all isolates. Terbinafine had the lowest MIC range of 0.001 g/ml to 0.09 g/ml and MIC50 was low at 0.005 g/ml and MIC90 was also low at 0.04 g/ml against T.rubrum; and MIC range of 0.001μg/ml to 0.19μg/ml with a MIC50 of 0.01μg/ml and MIC90 at 0.09μg/ml against T.mentagrophytes. Itraconazole showed antifungal activity superior to that of fluconazole, with a MIC range of 0.04g/ml to 1.56g/ml, with MIC50 at 0.19μg/ml and MIC90 at 1.56g/ml against T.rubrum; and MIC range of 0.04μg/ml to 1.56μg/ml, with MIC50 at 0.19μg/ml and MIC90 at 0.78μg/ml against T.mentagrophytes. Griseofulvin appears to be still a potent drug for management of dermatophytoses. Griseofulvin had a MIC range of 0.15g/ml to 5.07 g/ml with MIC50 at1.26 g/ml and MIC90 at 2.53 g/ml against T.rubrum; and MIC range of 0.31μg/ml to 5.07μg/ml with MIC50 at 1.26μg/ml and MIC90 at 2.53μg/ml against T.mentagrophytes. Fluconazole showed a high MIC range of 0.19 g/ml to 50 g/ml and MIC50 was high at 1.56g/ml and MIC90 was also high at 12.5 g/ml against T.rubrum; and a high MIC range of 0.09μg/ml to 25.0μg/ml, with MIC50 at 1.56μg/ml and MIC90 at 12.5μg/ml towards T.mentagrophytes. The technique was found to be easy to perform and reliable with consistent results.

In vitro antifungal susceptibility of griseofulvin, fluconazole, itraconazole and terbinafine against clinical isolates of Trichophyton rubrum and Trichophyton mentagrophytes

2014 ◽  
Vol 1 (4) ◽  
pp. 26-28
Author(s):  
KR Reddy ◽  
SR Reddy
Keyword(s):  
Trichophyton Rubrum ◽  
Clinical Laboratory ◽  
Antifungal Susceptibility ◽  
Trichophyton Mentagrophytes ◽  
Drug Susceptibility ◽  
Antifungal Drugs ◽  
Clinical Isolates ◽  
National Committee ◽  
In Vitro Antifungal Susceptibility

Investigations on antifungal drug susceptibility were carried out on 90 clinical isolates of Trichophyton rubrum, and Trichophyton mentagrophytes with four antifungal drugs, namely griseofulvin, fluconazole, itraconazole and terbinafine as suggested by National Committee for Clinical Laboratory Standards (NCCLS) M27A (1997) document by broth macrodilution method to standardize in vitro antifungal susceptibility testing and to find out the Minimum Inhibitory Concentration (MIC) of the drugs. In this study, terbinafine was found to be the most efficient drug for all isolates. Terbinafine had the lowest MIC range of 0.001 g/mlto 0.09 g/ml and MIC50 was low at 0.005 g/ml and MIC90 was also low at 0.04 g/ml against T. rubrum; and MIC range of 0.001pg/ml to 0.19pg/ml with a MIC50 of 0.01pg/ml and MIC90 at 0.09 pg/ml against T. mentagrophytes. Itraconazole showed antifungal activity superior to that of fluconazole, with a MIC range of 0.04g/ml to l.56g/ml, with MIC50 at 0.19pg/m land MIC90 at l.56g/ml against T. rubrum; and MIC range of 0.04.g/ml to 1.56pg/ml, with MIC50 at 0.19pg/ml and MIC90 at 0.78pg/ml against T. mentagrophytes. Griseofulvin appears to be still a potent drug for management of dermatophytoses. Griseofulvin had a MIC range of 0.15g/ml to 5.07 g/ml with MIC50 at l.26 g/ml and MIC90 at 2.53 g/ml against T. rubrum; and MIC range of 0.31pg/ml to 5.07pg/ml with MIC50 at 1.26pg/ml and MIC90 at 2.53pg/ml against T. mentagrophytes. Fluconazole showed a high MIC range of 0.19 g/ml to 50 g/ml and MIC50 was high at 1.56g/ml and MIC90 was also high at 12.5 g/ml against T. rubrum; and a high MIC range of 0.09pg/ml to 25.0pg/ml, with MIC50 at 1.56pg/ml and MIC90 at 12.5pg/ml towards T. mentagrophytes. The technique was found to be easy to perform and reliable with consistent results.DOI: http://dx.doi.org/10.3126/jucms.v1i4.9569 Journal of Universal College of Medical Sciences (2013) Vol.1 No.04: 26-28

scholarly journals Quantitation of Candida albicans Ergosterol Content Improves the Correlation between In Vitro Antifungal Susceptibility Test Results and In Vivo Outcome after Fluconazole Treatment in a Murine Model of Invasive Candidiasis

2000 ◽  
Vol 44 (8) ◽  
pp. 2081-2085 ◽  
Author(s):  
Beth A. Arthington-Skaggs ◽  
David W. Warnock ◽  
Christine J. Morrison
Keyword(s):  
Murine Model ◽  
Invasive Candidiasis ◽  
Clinical Laboratory ◽  
Antifungal Susceptibility ◽  
Resistant Isolate ◽  
National Committee ◽  
Ergosterol Content ◽  
In Vitro Antifungal Susceptibility

ABSTRACT MIC end point determination for the most commonly prescribed azole antifungal drug, fluconazole, can be complicated by “trailing” growth of the organism during susceptibility testing by the National Committee for Clinical Laboratory Standards approved M27-A broth macrodilution method and its modified broth microdilution format. To address this problem, we previously developed the sterol quantitation method (SQM) for in vitro determination of fluconazole susceptibility, which measures cellular ergosterol content rather than growth inhibition after exposure to fluconazole. To determine if SQM MICs of fluconazole correlated better with in vivo outcome than M27-A MICs, we used a murine model of invasive candidiasis and analyzed the capacity of fluconazole to treat infections caused by C. albicansisolates which were trailers (M27-A MICs at 24 and 48 h, ≤1.0 and ≥64 μg/ml, respectively; SQM MIC, ≤1.0 μg/ml), as well as those which were fluconazole sensitive (M27-A and SQM MIC, ≤1.0 μg/ml) and fluconazole resistant (M27-A MIC, ≥64 μg/ml; SQM MIC, 54 μg/ml). Compared with the untreated controls, fluconazole therapy increased the survival of mice infected with a sensitive isolate and both trailing isolates but did not increase the survival of mice infected with a resistant isolate. These results indicate that the SQM is more predictive of in vivo outcome than the M27-A method for isolates that give unclear MIC end points due to trailing growth in fluconazole.

scholarly journals In vitro susceptibilities of clinical and environmental isolates of Cryptococcus neoformans to five antifungal drugs.

1996 ◽  
Vol 40 (3) ◽  
pp. 822-824 ◽  
Author(s):  
S P Franzot ◽  
J S Hamdan
Keyword(s):  
Cryptococcus Neoformans ◽  
Amphotericin B ◽  
Clinical Laboratory ◽  
Antifungal Drugs ◽  
Clinical Isolates ◽  
National Committee ◽  
Environmental Isolates ◽  
Susceptibility Data

A total of 53 Cryptococcus neoformans strains, including clinical and environmental Brazilian isolates, were tested for their susceptibilities to amphotericin B, 5-flucytosine, ketoconazole, fluconazole, and itraconazole. The tests were performed according to the National Committee of Clinical Laboratory Standards recommendations (document M27-P). In general, there was a remarkable homogeneity of results for all strains, and comparable MICs were found for environmental and clinical isolates. This paper represents the first contribution in which susceptibility data for Brazilian C. neoformans isolates are provided.

scholarly journals In Vitro Antifungal Susceptibility and Molecular Characterization of Clinical Isolates of Fusarium verticillioides (F. moniliforme) and Fusarium thapsinum

2008 ◽  
Vol 52 (6) ◽  
pp. 2228-2231 ◽  
Author(s):  
Mónica Azor ◽  
Josepa Gené ◽  
Josep Cano ◽  
Deanna A. Sutton ◽  
Annette W. Fothergill ◽  
...  
Keyword(s):  
Active Drug ◽  
Fusarium Verticillioides ◽  
Antifungal Susceptibility ◽  
Antifungal Drugs ◽  
Clinical Isolates ◽  
Microdilution Method ◽  
Fusarium Thapsinum ◽  
In Vitro Antifungal Susceptibility

ABSTRACT A microdilution method was used to test 11 antifungal drugs against clinical isolates of Fusarium thapsinum and three different phylogenetic clades of Fusarium verticillioides that were characterized by sequencing a region of the β-tubulin gene. Terbinafine was the most-active drug against both species, followed by posaconazole against F. verticillioides.

scholarly journals The Trailing End Point Phenotype in Antifungal Susceptibility Testing Is pH Dependent

1999 ◽  
Vol 43 (6) ◽  
pp. 1383-1386 ◽  
Author(s):  
Kieren A. Marr ◽  
Tige R. Rustad ◽  
John H. Rex ◽  
Theodore C. White
Keyword(s):  
Minimal Medium ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
Ph Effect ◽  
Antifungal Susceptibility ◽  
Drug Susceptibility ◽  
Drug Susceptibility Testing ◽  
National Committee ◽  
End Points

ABSTRACT The interpretation of end points in azole antifungal drug susceptibility testing is problematic, in part due to incomplete growth inhibition of Candida species. Such trailing growth can cause the MICs of fluconazole for some isolates to be low (<1 μg/ml) after 24 h of growth but much higher (>64 μg/ml) after 48 h. Isolates having this type of growth have been described as having a low-high phenotype. Although these isolates would be considered resistant by current National Committee of Clinical Laboratory Standards definitions, growing evidence suggests that they are susceptible in vivo. To further characterize these isolates in vitro, microdilution susceptibility testing comparing the complex defined medium RPMI 1640 to a defined minimal medium (yeast nitrogen broth) was performed. Isolates having trailing growth in MOPS (morpholinepropanesulfonic acid)-buffered RPMI 1640 (pH 7.0) were found to have clear end points in the minimal medium at its native pH of 4.5. The pH of the medium influenced the low-high phenotype, as these same isolates trailed in minimal medium adjusted to a pH of ≥6.0 but did not trail in RPMI 1640 adjusted to a pH of ≤5.0. This pH effect was independent of the medium buffering capacity, as trailing was decreased in both minimal medium and RPMI 1640 (pH 4.5) buffered in citrate. Adjustment in the pH of MOPS-buffered RPMI 1640 reduced trailing in multiple strains of Candida albicans without affecting the MICs for isolates having known susceptible (low-low) and resistant (high-high) phenotypes. Adjustment of the medium pH could be considered to eliminate trailing in azole drug susceptibility testing.

scholarly journals Population differentiation, antifungal susceptibility, and host range of Trichophyton mentagrophytes isolates causing recalcitrant infections in humans and animals

2020 ◽  
Vol 39 (11) ◽  
pp. 2099-2113
Author(s):  
Sebastian Gnat ◽  
Dominik Łagowski ◽  
Aneta Nowakiewicz ◽  
Marcelina Osińska ◽  
Łukasz Kopiński
Keyword(s):  
Host Range ◽  
Susceptibility Testing ◽  
Antifungal Susceptibility ◽  
Trichophyton Mentagrophytes ◽  
Antifungal Susceptibility Testing ◽  
Antifungal Drugs ◽  
Genomic Diversity ◽  
Clinical Isolates ◽  
Chemical Groups

Abstract The major problems in determining the causative factors of the high prevalence of dermatophytoses include the lack of a well-standardized antifungal susceptibility testing method, the low consistency of in vitro and clinical minimal inhibitory concentration values, the high genomic diversity of the population, and the unclear mechanism of pathogenicity. These factors are of particular importance when the disease is recalcitrant and relapses. Herein, we identified and characterized Trichophyton mentagrophytes isolates obtained from therapy-resistant cases in humans and animals. We used genomic diversity analysis of 17 human and 27 animal clinical isolates with the MP-PCR technique, determined their phenotypic enzymatic activity and host range, and performed antifungal susceptibility testing to currently available antifungal drugs from various chemical groups. Genomic diversity values of 35.3% and 33.3% were obtained for clinical isolates from humans and animals, respectively, yet without any relationship to the host species or antifungal drug to which resistance in therapy was revealed. The highest activity of keratinase enzymes was recorded for fox, guinea pig, and human hairs. These hosts can be considered as the main species in the host range of these isolates. A phenyl morpholine derivative, i.e. amorolfine, exhibited superior activity against strains obtained from both humans and animals with the lowest MIC50. Interestingly, high compliance of terbinafine in vitro resistance with clinical problems in the treatment with this substance was shown as well. The high resistance of dermatophytes to drugs is the main cause of the recalcitrance of the infection, whereas the other features of the fungus are less important.

Use of spectrophotometric reading for in vitro antifungal susceptibility testing ofAspergillusspp.

1999 ◽  
Vol 45 (10) ◽  
pp. 871-874 ◽  
Author(s):  
Eric Dannaoui ◽  
Florence Persat ◽  
Marie-France Monier ◽  
Elisabeth Borel ◽  
Marie-Antoinette Piens ◽  
...  
Keyword(s):  
Amphotericin B ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
Antifungal Susceptibility ◽  
Reference Method ◽  
Antifungal Susceptibility Testing ◽  
Aspergillus Species ◽  
National Committee ◽  
Broth Microdilution Method

A comparative study of visual and spectrophotometric MIC endpoint determinations for antifungal susceptibility testing of Aspergillus species was performed. A broth microdilution method adapted from the National Committee for Clinical Laboratory Standards (NCCLS) was used for susceptibility testing of 180 clinical isolates of Aspergillus species against amphotericin B and itraconazole. MICs were determined visually and spectrophotometrically at 490 nm after 24, 48, and 72h of incubation, and MIC pairs were compared. The agreement between the two methods was 99% for amphotericin B and ranged from 95 to 98% for itraconazole. It is concluded that spectrophotometric MIC endpoint determination is a valuable alternative to the visual reference method for susceptibility testing of Aspergillus species.Key words: antifungal, susceptibility testing, Aspergillus, spectrophotometric reading.

scholarly journals Comparison of NCCLS and 3-(4,5-Dimethyl-2-Thiazyl)-2,5-Diphenyl-2H-Tetrazolium Bromide (MTT) Methods of In Vitro Susceptibility Testing of Filamentous Fungi and Development of a New Simplified Method

2000 ◽  
Vol 38 (8) ◽  
pp. 2949-2954 ◽  
Author(s):  
Joseph Meletiadis ◽  
Jacques F. G. M. Meis ◽  
Johan W. Mouton ◽  
J. Peter Donnelly ◽  
Paul E. Verweij
Keyword(s):  
Filamentous Fungi ◽  
Clinical Laboratory ◽  
Colorimetric Method ◽  
Antifungal Drugs ◽  
National Committee ◽  
In Vitro Susceptibility ◽  
Initial Inoculum ◽  
Tetrazolium Bromide ◽  
Mtt Method

The susceptibility of 30 clinical isolates belonging to six different species of filamentous fungi (Aspergillus fumigatus, Aspergillus flavus, Scedosporium prolificans, Scedosporium apiospermum, Fusarium solani, and Fusarium oxysporum) was tested against six antifungal drugs (miconazole, voriconazole, itraconazole, UR9825, terbinafine, and amphotericin B) with the microdilution method recommended by the National Committee for Clinical Laboratory Standards (NCCLS) (M38-P). The MICs were compared with the MICs obtained by a colorimetric method measuring the reduction of the dye 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) to formazan by viable fungi. The levels of agreement between the two methods were 96 and 92% for MIC-0 (clear wells) and MIC-1 (75% growth reduction), respectively. The levels of agreement were always higher for Aspergillus spp. (97% ± 2.5%), followed byScedosporium spp. (87% ± 10.3%) and Fusariumspp. (78% ± 7.8%). The NCCLS method was more reproducible than the MTT method: 98 versus 95% for MIC-0 and 97 versus 90% for MIC-1. However, the percentage of hyphal growth as determined visually by the NCCLS method showed several discrepancies when they were compared with the percentages of MTT reduction. A new simplified assay that incorporates the dye MTT with the initial inoculum and in which the fungi are incubated with the dye for 48 h or more was developed, showing comparable levels of agreement and reproducibility with the other two methods. Furthermore, the new assay was easier to perform and more sensitive than the MTT method.

scholarly journals In Vitro Amphotericin B Resistance in Clinical Isolates of Aspergillus terreus, with a Head-to-Head Comparison to Voriconazole

1999 ◽  
Vol 37 (7) ◽  
pp. 2343-2345 ◽  
Author(s):  
Deanna A. Sutton ◽  
Stephen E. Sanche ◽  
Sanjay G. Revankar ◽  
Annette W. Fothergill ◽  
Michael G. Rinaldi
Keyword(s):  
Amphotericin B ◽  
Aspergillus Terreus ◽  
Clinical Laboratory ◽  
Immunocompromised Host ◽  
Clinical Isolates ◽  
National Committee ◽  
High Dose ◽  
Etiologic Agents ◽  
Disseminated Disease

Amphotericin B therapy continues to be the “gold standard” in the treatment of invasive aspergillosis in the immunocompromised host. Although Aspergillus fumigatus and Aspergillus flavus constitute the major species, several reports have described invasive pulmonary or disseminated disease due to the less common Aspergillus terreus and dismal clinical outcomes with high-dose amphotericin B. We therefore evaluated 101 clinical isolates of A. terreus for their susceptibility to amphotericin B and the investigational triazole voriconazole by using the National Committee for Clinical Laboratory Standards M27-A method modified for mould testing. Forty-eight-hour MICs indicated 98 and 0% resistance to amphotericin B and voriconazole, respectively. We conclude that A. terreus should be added to the list of etiologic agents refractory to conventional amphotericin B therapy and suggest the potential clinical utility of voriconazole in aspergillosis due to this species.

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