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Volume 25 |
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Number 2 |
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Published by : PROFESSIONAL MEDICAL PUBLICATIONS |
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Volume 25 |
April - June 2009 (Part-I) |
Number 2 |
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In vitro interactions of miconazole with
sulfametoxazole against candida species
Majid Zarrin1, Mahmoud Jamshidian2, Mohammad Jafari3
ABSTRACT
Objective: Finding the synergism effect of miconazole and sulfametoxazole on 4 species of candida.
Methodology: Interactions of miconazole in combination with sulfametoxazole were tested in buffered yeast-nitrogen base using checkerboard method. Plates were inoculated with 20µl cells suspensions of each yeast and incubated at 30°C for 24 hour. In this method, the MICs were described as the lowest antimicrobial concentration inhibiting visible fungal growth on the plates. Minimal fungicidal concentration (MFC) was described as the first tube showing no growth on the plate.
Results: The MIC of miconazole was 11.30µg/ml for Candida albicans. 8.9µg/ml for Candida tropicalis 8.6µg/ml for Candida papapsilosis and 5.2µg/ml for Candida krusei. When miconazole were combined with sulfametoxazole, miconazole MICs decreased to 1.54µg/ml for C. albicans, 0.5µg/ml for C. tropicalis, 0.383µg/ml for C. parapsilosis and 0.275µg/ml for C. krusei.
Conclusion: The data show that combination of miconazole and sulfametoxazole was synergistic on C. albicans, C. tropicalis, C. parapsilosis and C. krusei.
KEY WORDS: Candida, Miconazole, Sulfametoxazole, Synergism.
Pak J Med Sci April - June 2009 Vol. 25 No. 2 243-246
How to cite this article:
Zarrin M, Jamshidian M, Jafari M. In vitro interactions of miconazole with sulfametoxazole against candida species. Pak J Med Sci 2009;25(2):243-246.
1. Majid Zarrin,
Department of Parasitology and Mycology,
Medical School,
Jundishapur University of Medical Sciences,
Ahwaz, Iran.
2. Mahmoud Jamshidian,
Department of Pathobiology,
School of Veterinary Medicine,
Shahid Chamran University, Ahwaz, Iran.
3. Mohammad Jafari,
Department of Parasitology and Mycology,
Medical School,
Jundishapur University of Medical Sciences,
Ahwaz, Iran.
Correspondence
Dr. Majid Zarrin,
Department of Parasitology and Mycology,
Medical School,
Jundishapur University of Medical Sciences,
Ahwaz, Iran.
Email: mjzarrin@yahoo.co.uk
* Received for Publication: July 5, 2008
* Revision Accepted: February 7, 2009
INTRODUCTION
Candida species, generally C. albicans, are the most common causative agents of human fungal infections.
1,2 The increase in the rate of opportunist diseases with Candida has resulted from the growing use of broadspectrum antibacterial drugs, and from an increasing number of immunocompromised patients because of the use of aggressive chemotherapy for cancer and of immunosuppressive drugs in organ tranplation. The progress of infection by the human immunodeficiency virus (HIV) is another significant reason for immunocompromise.3 Antifungal agents are used increasingly both as curative and prophylactic agents,4 which, in turn, has led to the widespread appearance of resistant strains.4,5Opportunistic fungal infections have increased dramatically in recent years as a result of increased immunisuppression associated with AIDS, organ and bone marrow transplantation, and aggressive treatment of cancer and autoimmune disorders. Miconazole, (MIC), an antifungal imidazole, has been used world-wide in the treatment of superficial mycoses. Many clinicians believe the regional treatment with MIC is a relatively effective in the treatment of most mycotic infections.
6,7Systemic (interavenous) use of miconazole for invasive fungal infections had initial positive results
7 but was discontinued due to toxicity of the vehicle and reports of hepatic and cardiovascular side effects.8,9 Combination therapy might be a promising approach in such circumstances. The use of antifungal combinations may increase the rates of microbial killing, shorten the durations of therapy, avoid the emergence of drug resistance, and expand the spectrum of activity.Therefore, in this study, we investigated the in vitro activities of miconazole, alone and in combination with sulfametoxazole, against C. albicans, C. tropicalis, C. parapsilosis and C. krusei.
METHODOLOGY
Organisms: Strains of C. albicans, C. tropicalis, C. parapsilosis and C. Krusei collected by the medical mycology laboratory at the Ahwaz Jundishapur University of Medical Sciences. Cultures of C. albicans, C. tropicalis, C. parapsilosis and C. krusei were grown on Sabouraud’s dextrose agar at room temperature for three days. Each isolate was suspended in 5 ml of sterile distilled water including tween 80 and vortexed thoroughly to achieve a smooth suspension. Final suspensions were adjusted to 90% transmittance at 530 nm, corresponding to 10
6 CFU/m.Drug solutions:
Stock solutions of miconazole (Behvazan) and sulfametoxazole (Rooz Daru) were solved in dimethy sulfoxide. These solutions were kept at room temperature for 30 minutes for self-sterilisation. The drug solutions were subsequently stored at -70°C for further experiences.Method: The susceptibility testing of miconazole and sulfametoxazole were performed in yeast nitrogen base glucose (YNBG) agar buffered with phosphate buffer (pH 7). In the first step both miconazole with sulfametoxazole were tested separately. Final drug concentrations for miconazole was 64, 32, 16, 14, 12, 10, 8, 6, 4, 2, 1, 0.5, 0.25, 0.125µg/ml. Final drug concentrations for sulfametoxazole was 200,100, 50, 25, 12.5µg/ml. Combination rows each contained miconazole in dilutions from 64 to 0.125µg/ml with sulfametoxazole from 200 to 12.5µg/ml. Drug activity was assessed by a checkerboard method. Growth control tube contained media with and without each of the drugs and dimethy sulfoxide in plates. Plates were inoculated with 20µl yeast suspensions of each organism and incubated at 30°C for 24 hour. In this method, the MICs were described as the lowest antimicrobial concentration inhibiting visible fungal growth on the plates. Minimal fungicidal concentration (MFC) was described as the first tube showing no growth on the plate.
RESULTS
The checkerboard method was performed in this study to investigate the interaction between miconazole and sulfametoxazole against isolates of C. albicans, C. tropicalis, C. parapsilosis and C. krusei. The susceptibility results achieved for the isolates are shown in Table-I.
All isolates were tested against both miconazole and sulfametoxazole alone and in combination with each other. In this study, 25 isolates of C. albicans, 20 isolates of C. tropicalis, 15 isolates of C. parapsilosis and 10 isolates of C. krusei were tested. Miconazole MIC means was 11.36, 8.9, 8.6 and 5.2 µg/ml for C. albicans, C. tropicalis, C. parapsilosis and C. krusei respectively (Table-I).
The addition of sulfametoxazole to miconazole gave variable results. When miconazole was combined with sulfametoxazole, there were significant reductions in concentration of miconazole. Synergy was demonstrated against C. albicans, C. tropicalis, C. parapsilosis and C. krusei. In all isolates, the concentration of miconazole in combination with sulfametoxazole was reduced.
Table-I shows a list the MIC at which 100% of the strains were inhibited by miconazole and miconazole + sulfametoxazole. When inhibitory concentration of miconazole were tested with 12.5µg/ml sulfametoxazole, miconazole MICs decreased to 1.54, 0.5, 0.383 and 0.275µg/ml in C. albicans, C. tropicalis, C. parapsilosis and C. krusei respectively.
Sulfametoxazole alone also was used against all isolates tested. When subinhibitory concentrations of miconazole were used, sulfametoxazole MICs decreased significantly in isolates. Antagonism was not noted with these drug combinations against any of the isolates tested.
DISCUSSION
Certain gaps in the therapeutic efficiency of antifungal drugs may be a necessary trade-off for the benefits of lower toxicity
10. In vitro susceptibility test results can afford a helpful guide for selection of antifungal drug therapy, particularly when reliable interpretive breakpoints are used. They are particularly valuable in identifying yeast isolates that are unlikely to respond to a given antifungal agent. Nevertheless, an isolate’s in vitro susceptibility to a certain drug does not predict the in vivo efficacy of that drug or, more significantly, the therapeutic outcome, which is often influenced by numerous factors, including drug interactions, host factors and pharmacokinetics11. Standardized methods for culture and in vitro antifungal susceptibility testing with high intra-laboratory and inter-laboratory reproducibility are also able to afford reliable information for identifying local patterns, which must be taken into account in the selection of emperical therapy, mainly in cases of severe candidiasis or candidema.12,13The need for novel antifungal regimens promoted this study of the synergic effect of sulfametoxazole in association with miconazole. sulfametoxazole was found to have a marked effect of C. albicans and non-albicans.
Delineation of drug susceptibilities of Candida species or strains to antifungal drugs has become of increased importance because of the identification of multi-azole-resistant strains and the emergence of species other than C. albicans as pathogen.
14-16 The lack of efficient antifungal therapy encouraged us to test the candida isolates against combinations of drugs that have shown activity against other fungi. By establishing dose-response based on fungal cell suspensions, we were able to confirm our finding by the checkerboard method. The results suggest that MIC strains were relatively high to miconazole and the concentration of the miconazole was decreased significantly when sulfametoxazole was added in the media. The lowest MIC was 0.275µg/ml for C. krusei and the highest was 1.54 for C. albicans. Based on our data the addition of sulfametoxazole to miconazole is expected to have enhanced activity against selected isolates. This research demonstrates that susceptibility tests in vitro may be important predictors of the most suitable drug or drugs to use clinically against Candida species.REFERENCES
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12. Pfaller MA, Diekema DJ, Jones RN, Messer SA, Hollis RJ, SENTRY participants group. Trends in antifungal susceptibility of Candida spp. isolated from pediatric and adults patients with bloodstream infections: SENTRY antimicrobial surveillance program, 1997 to 2000. J Clin Microbiol 2002;40:852-6.
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14. Galgiani JN, Lewis ML. In vitro studies of the activities of the antifungal triazoles SCH56592 and itraconazole against Candida albicans, Cryptococcus neoformans, and other pathogenic yeasts. Antimicrob Agents Chemother 1997;41:180-3.
15. Pfaller MA, Messer S, Jones RN. Activity of a new triazole, Sch 56592, compared with those of four other antifungal agents tested against clinical isolates of Candida spp. and Saccharomyces cerevisiae. Antimicrob Agents Chemother 1997;41:233-5.
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