ACRIDINE ORANGE FLUOROCHROMING AND ULTRAVIOLET ABSORPTION OF STAPHYLOCOCCUS AUREUS CELLS MODIFIED BY UNBALANCED NUCLEIC ACID AND CELL WALL SYNTHESIS

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Abstract
Cultures of Staphylococcus aureus were grown in the presence of five different antimetabolites (5-fluorodeoxyuridine, aminopterin, 8-azaguanine, mitomycm-C, 5-fluorouracil) active against ceil walls and (or) nucleic acids. Fluorescence microscopy of smears stained with acridine orange revealed reddish and green cells in both treated and untreated cultures. There were less than 20% of reddish cells in untreated cultures and more than 40% in treated cultures. Treated cultures contained fewer viable organisms. All antimetabolites except mitomycin-C produced a diminution in the nucleic acids, chemically determined as percentage of dry weight of bacteria. Only 5-fluorouracil increased the RNA/DNA ratio. However, with ultraviolet microscopy at 260 mμ wavelength the absorption of reddish cells is much higher than that of the green cells, which, at the cellular level, seemed to indicate a greater nucleic acid content. With ultraviolet or with fluorescence microscopy we have obtained similar evidence of the cellular heterogeneity produced by antimetabolites in bacterial populations.