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Synergistic Effects of Efflux Pump Modulators on the Azole Antifungal Susceptibility of Microsporum canis

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Abstract

The microbiologic and clinical resistance of dermatophytes is seldom reported, and the mechanisms associated with resistance are not well known. This study investigated the effect of efflux pump modulators (EPMs) (i.e., haloperidol HAL and promethazine PTZ) and their inhibiting activity on the minimum inhibitory concentrations of itraconazole (ITZ) and fluconazole (FLZ) against selected M. canis strains. M. canis strains with low (≤ 1 μg/ml itraconazole and < 64 μg/ml fluconazole) and high (> 1 μg/ml itraconazole and ≥ 64 μg/ml fluconazole) azole MIC values were tested using Checkerboard microdilution assay. The disk diffusion assay, the minimum fungicidal concentration and the time-kill assay were also performed in order to confirm the results of checkerboard microdilution assay. The MIC values of ITZ and FLZ of M. canis decreased in the presence of subinhibitory concentrations of HAL and PTZ, the latter being more effective with a greater increased susceptibility. Synergism was observed in all strains with high azole MICs (FICI < 0.5) and no synergism in the strains with low azole MICs. A fungicidal activity was observed after 48 h of incubation when ITZ and FLZ were tested in combination with HAL or PTZ. These results suggest that the drug efflux pumps are involved in the defense mechanisms to azole drugs in M. canis strains. The synergism might be related to an increased expression of efflux pump genes, eventually resulting in azole resistance phenomena. Complementary studies on M. canis resistance are advocated in order to investigate the molecular mechanisms of this phenomenon.

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Funding

This research received no external funding.

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Authors and Affiliations

  1. Dipartimento Di Medicina Veterinaria, Università Degli Studi “Aldo Moro”, 70010, Valenzano (Bari), Italy

    Chioma Inyang Aneke, Wafa Rhimi, Domenico Otranto & Claudia Cafarchia

  2. Department of Veterinary Pathology and Microbiology, University of Nigeria, Nsukka, 410001, Nigeria

    Chioma Inyang Aneke

  3. Faculté Des Sciences de Bizerte, Université de Carthage, Zarzouna, Tunisia

    Wafa Rhimi

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  1. Chioma Inyang Aneke
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  2. Wafa Rhimi
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  3. Domenico Otranto
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  4. Claudia Cafarchia
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Contributions

Aneke Chioma Inyang and Cafarchia Claudia conceptualized the study and wrote the manuscript. Aneke Chioma Inyang and Rhimi Wafa performed the research. Aneke Chioma Inyang, Cafarchia Claudia and Otranto Domenico contributed to analysis and interpretation of the study data. Cafarchia Claudia and Otranto Domenico revised, edited and made intellectual inputs in the manuscript. All authors read and approved the final manuscript.

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Correspondence to Claudia Cafarchia.

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Handling editor: Nilce M. Martinez-Rossi.

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Aneke, C.I., Rhimi, W., Otranto, D. et al. Synergistic Effects of Efflux Pump Modulators on the Azole Antifungal Susceptibility of Microsporum canis. Mycopathologia 185, 279–288 (2020). https://doi.org/10.1007/s11046-019-00419-7

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  • DOI: https://doi.org/10.1007/s11046-019-00419-7

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