Ultrastructure of Candida ingens: a yeast that can assimilate volatile fatty acids.

A defined liquid medium was developed for culture of Candida ingens on either volatile fatty acids or glucose as carbon source. Buffering capacity at pH 5.4 was achieved by inclusion of 50 mM phthalic acid which is not assimilated. The yeast grew on C2-C5 acids, with lags in some cases that were dependent on the cultural history of the inoculum. The cells were notably large for Candida species, or yeasts in general, and were significantly more elongated on glucose substrate compared with butyrate. Traditional aldehyde fixatives and heavy metal staining yielded bland micrographs. However, the periodic acid-thiocarbohydrazide-silver proteinate stain of Thiéry resulted in informative, high contrast electron images for this species. The ultrastructure of the cell envelope was not greatly affected by carbon source or by growth habit except that a slime layer was more prevalent in pellicular cells compared with those grown in submerged culture. The slime layer was apparently stabilized by colloidal iron staining. The other feature of the cell envelope was a microfibrillar zone containing periodic acid reactive constituents. Subcellular organelles were typical of yeast species although there was a propensity for large vacuoles in pellicular cells. When cells were grown on fatty acids there was an increase in the number of microbodies compared with that observed on glucose. Microbodies were best demonstrated by diaminobenzidine-hydrogen peroxide staining of protoplasts, which were generated by dissolution of cell walls with snail digestive enzymes.