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SUMMARY: Wet and dry Gram staining procedures have different characteristics in respect to their rates of decolorization. It has been proposed that this difference is due to a Gram-positive substrate which dissociates (becomes Gram-negative) in the presence of water. However, the differences between wet and dry Gram procedures can also be explained on the hypothesis that water influences the rate of solvent permeation through cell envelopes. Since no direct proof exists for the reality of the proposed Gram-positive substrate, and since evidence is presented here which cannot be explained by such a hypothesis, it is felt that the solvent permeation concept should be considered seriously as an explanation for the decolorization differences observed between wet and dry Gram procedures.

The Effect of Moisture on Gram Differentiation, and its Relation to Proposed Gram-Positive Substrates in Yeast and Other Organisms