Metconazole

AFST with 14-α demethylation inhibiting fungicides (DMIs) was performed following the EUCAST methodology as described before. Environmental DMIs tested for microdilution assay were two imidazole drugs, imazalil and prochloraz; and five triazole drugs, tebuconazole, epoxiconazole, difenoconazole, bromuconazole and metconazole, all of them purchased at Sigma-Aldrich Química, Madrid, Spain. The final concentrations of each azole drug tested ranged from 0.06 to 32 mg/L. MICs were visually read as described before. Clinical breakpoints for DMIs have not been standardized yet so wild-type strain MICs were considered as susceptible.

Seventeen agricultural fungicides (test range 32–0.03 mg/L), five azole fungicide metabolites (test range 16–0.016 mg/L) and four medical azoles were included (test range 32–0.03 mg/L). The agricultural fungicides (bixafen, boscalid, fluxapyroxad, fluopyram, folpet, azoxystrobin, prothioconazole, paclobutrazole, epoxiconazole, propiconazole, tebuconazole, difenoconazole, metconazole, prothiozonacole-desthio, prochloraz, imazalil were purchased from Sigma-Aldrich, Søborg, Denmark, whereas mefentrifluconazol was purchased from LGC Standards (Teddington, Middlesex, United Kingdom www.lgcstandards.com, accessed on 26 January 2021). The metabolites included: 1,2,3-triazole (Sigma-Aldrich, Søborg, Denmark), 1,2,4-triazole and triazole sulfonamide (Toronto Research Chemicals, Toronto, Canada), and triazole alanine and triazole acetate (HPC Standards GmbH, Borsdorf, Germany). The medical antifungal agents used were itraconazole, voriconazole, isavuconazole and posaconazole (all from Sigma-Aldrich, Søborg, Denmark, except isavuconazole, which derived from Basilea).