Fungitoxicity of muramidase. Ultrastructural damage to Candida albicans.

The antifungal activity of hen egg-white lysozyme was investigated in vitro using a hypotonic medium designed to support an anabolic cellular state with minimal growth stimulation. The pathogenic yeast Candida albicans was found to be sensitive to microgram amounts of hen egg-white lysozyme. This susceptibility was evidenced by sluggish growth and a dose-dependent killing process. Transmitted and scanning electron microscopic observations on lysozyme-treated C. albicans yeast cells revealed the following ultrastructural modifications: (1) plasmolysis, vacuolar expansion and wrinkled surface configuration; (2) unremitting accumulation of wall-like material that bulged into the periplasmic space; (3) qualitative changes in the organization of the wall. Ongoing structural modifications within the wall were highlighted with the cationic heavy metal dye ruthenium red. A disruption in the permeability of the cytoplasmic membrane was evidenced by modification 1 and by differential staining characteristics in light microscopy. The superimposed osmotic imbalance was identified as the cause of cell death. It is proposed that lysozyme acts on C. albicans by two distinct complementary mechanisms: enzymatic hydrolysis of N-glycosidic bonds that link polysaccharides and structural proteins of the wall; injury to the cytoplasmic membrane as a result of a cationic protein kind of interaction.