This work evaluated the effect of adjunct autochthonous cultures on the chemical, microbiological and sensory characteristics of Fiore Sardo cheese during ripening. A total of twelve batches of cheeses were manufactured according to the technical Disciplinary of Fiore Sardo cheese, with and without different combinations of autochthonous strains isolated from the native microflora of artisanal Fiore Sardo. There were no significant differences in the cheese compositional parameters between experimental and control cheeses, but the addition of cultures led to a statistically significant decrease in pH values in experimental cheeses. The evolution of total mesophilic bacteria, total coliforms and lactic acid bacteria were significantly influenced by the addition of autochthonous cultures in most of the experimental cheeses. As for sensory characteristics, all the experimental cheeses reported significantly higher scores especially for shape, texture, interior openings, taste and aftertaste. This study demonstrated the beneficial effect of the addition of selected autochthonous cultures in accelerating the disappearance of undesirable flora and improving the typical sensory characteristics of the cheese, and confirmed the importance of ewes' milk as a source of technologically interesting strains that could be used to ensure a higher quality of artisanal cheese productions.