We report on the study of surfactant-free silver nanoparticles synthesized using non-hydrolytic sol–gel methods for applications in straw bale constructions. Micro-organism infestation in green constructions is of concern as their proliferation tends to induce health problems. We demonstrate the biocidal properties of these Ag nanoparticles and their efficacy against fungi. Outdoor tests with Ag nanoparticles have demonstrated the effective protection of straw against micro-organisms. Indoor tests using broth liquid are compared with a method of testing we recently developed where the possible nature of the biocidal properties of the silver nanoparticles are further probed. In contrast to the commonly reported results, this study shows that Ag nanoparticles synthesized using non-hydrolytic sol–gel methods have antifungal properties against common fungi in outdoor conditions which demonstrate high potential in related applications.