Fluopyram

Synchronized L4 hermaphrodite worms (N2, RP2698, and RP2674) were individually placed on NGM plates seeded with E. coli OP50 and exposed to various concentrations of cyclobutrifluram or fluopyram (Sigma-Aldrich, Burlington, MA, USA). The worms were transferred to new plates every 24 h until egg laying stopped. The total number of eggs, adult hermaphrodites, and males were scored. The average brood size, embryonic viability, and incidence of males were determined. Statistical significance was assessed using a two-tailed Student’s t-test with Welch’s correction, p-value ≤ 0.05.

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).