Fifty-six dairy bacteria belonging to the genera Lactococcus, Lactobacillus, Pediococcus, Propionibacterium, Streptococcus, Enterococcus, Leuconostoc, and Brevibacterium were screened for antifungal activity against four species of fungi relevant to the cheese industry (Penicillium discolor, Penicillium commune, Penicillium roqueforti, and Aspergillus vesicolor). Most of the active strains belonged to the genus Lactobacillus, whereas Penicillium discolor was found to be the most sensitive of the four fungi investigated. Further studies on P. discolor showed antifungal activity only below pH 5. This effect of pH suggests that organic acids present in the culture could be involved in the detected activity. Determination of acid composition revealed lactic acid production for active dairy strains and the presence of acetic acid in active as well as inactive strains. It was demonstrated that the undissociated acetic acid originates from the bacterial growth medium. The synergistic effect of the acetic acid present and the lactic acid produced was likely the main factor responsible for the antifungal properties of the selected bacteria. These results could explain some discrepancies in reports of the antifungal properties of lactic acid bacteria, since the role of acetic acid has not been considered in previous studies.