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Background  Current drug regimens for cutaneous leishmaniasis (CL) include toxic systemic therapies such as amphotericin B (AB) and pentavalent antimonials. Fluconazole (FZ) is a well-tolerated potential oral alternative for the management CL. To date, few objective data exist to guide clinical decision-making when selecting a therapeutic agent a priori, and standardized, clinically-approved drug susceptibility testing platforms for  Leishmania  spp. have yet to be established. The Sensititre™ YeastOne™ YO9 plate is a commercialized drug susceptibility plate including AB and FZ used for routine testing of non-fastidious yeast. Our objective was to adapt the readily available Sensititre™ YeastOne™ YO9 plate, to determine drug susceptibility profiles of AB and FZ in cultured isolates of Old World and New World  Leishmania  spp. for the treatment of CL.    Methods  Promastigotes were cultured in Tobie’s medium with Locke’s overlay until log phase growth was achieved, inoculated into the Sensititre™ system, and incubated over 96 H . minimum  inhibitory concentrations (MICs) were determined colorimetrically, and promastigote death was assessed by conventional microscopy out to 96- h. Colour change correlated to MIC values.    Results  All strains tested exhibited MIC values for FZ that were ≥ 256 μg/mL. New World strains demonstrated reduced susceptibility to AB (0.25 μg/mL – 0.50 μg/mL AB) compared to Old World strains at 0.12 μg/mL AB ( p  = 0.02). Seventeen (61%) of 28  Viannia  isolates versus 82% (27/33) of non- Viannia  isolates were resistant at 0.12 μg/mL AB ( p  = 0.09). For  L. V. braziliensis  isolates, mean MIC for AB was 0.375 ± 0.14 μg/mL (range 0.25–0.50 μg/mL), while for isolates of  L. V. panamensis  it was 0.314 ± 0.26 μg/mL (range 0.12–1.0 μg/mL).    Conclusions  We adapted the Sensititre™ YeastOne™ YO9 plate for testing of  Leishmania  spp .  susceptibility profiles for commonly used antifungals in the treatment of CL, including AB and FZ. Given its current utility in mycology, optimization of the system for potential clinical implementation in parasitology should be pursued. However evaluation of clinically relevant amastigote-stage stages, and higher concentrations of FZ beyond the upper limit concentration of the Sensititre™ YeastOne™ Y09 plate would be required.

Susceptibility testing of Leishmania spp. against amphotericin B and fluconazole using the Sensititre™ YeastOne™ YO9 platform