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Dihydromyricetin-mediated silver nanoparticles (DMY-AgNPs) were synthesized and their efficacy against fungal pathogens tested in vitro. The shape of DMY-AgNPs appeared to be spherical with size of ~34 nm. Fourier-transform infrared (FT-IR) analysis indicated that -OH and C=O groups were involved in nanoparticle formation. The XRD pattern of DMY-AgNPs showed strong peaks at 38°, 44°, and 64°, corresponding to reflection from (111), (200), and (220) planes. Five opportunistic fungal pathogens, namely Aspergillus fumigatus, Aspergillus niger, Paecilomyces formosus, Candida albicans, and Candida parapsilosis, were isolated from patients suffering from respiratory tract infections. Growth of each fungal strain was inhibited by DMY-AgNPs. The zone of inhibition of DMY-AgNPs against A. fumigatus, A. niger, P. formosus, C. albicans, and C. parapsilosis was 17.6, 19.2, 22.2, 15.8, and 18.5 mm. The minimal inhibitory concentration was found to be 0.83, 0.73, 0.67, 0.95, and 0.89 µg mL−1, respectively. This is the first report on DMY-AgNPs as an effective antifungal agent. DMY-AgNPs are a potential alternative to commercially available antifungal fungicidals. [ABSTRACT FROM AUTHOR]