Silver particles are prepared by dewetting Ag films coated on glass using a fiber laser. The size of the particles is controlled in the range of 92 nm ~ 1.2 μm by adjusting the thickness of the Ag film. The structural properties and surface roughness of the particles are evaluated by means of scanning electron microscopy. In addition, the antifungal activity of the Ag particles is examined using spore suspensions of Colletotrichum gloeosporioides. It is shown that the particles with a size of 1.2 μm achieve 100% inhibition of the conidia growth of the Colletotrichum gloeosporioides after a contact time of just 5 min. Furthermore, the smaller particles also achieve a good antibacterial activity given a longer contact time. Similar results are observed in spore germination and pathogenicity tests performed on mango fruit and leaves. Overall, the results confirm that the Ag particles have an excellent antifungal effect on Colletotrichum gloeosporioides.