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Eucaryotic unicellular (a yeast, a cellular slime mold, and various protozoans) and two multicellular (aschelminths) microorganisms were saturated with gas at high pressures and rapidly decompressed. No effect was observed with pressures of argon up to 125 atm, nitrogen up to 175 atm, and helium up to 350 atm, showing that the induced gas supersaturations did not cause intracellular bubbles to form. With 25--50 atm higher gas pressures, the decompression usually produced killing and cell rupture, although differences in tolerances existed among the various organisms. Substantial fractions of the populations survived gas supersaturations well above the threshold values for massive spontaneous nucleation of bubbles in the water. When killing occurred, external rather than internal bubbles appeared to be the cause. Even with the 300 atm argon or nitrogen pressures, yeast cells were unaffected, apparently because of the external protection provided by their cell wall. It is concluded that the gas supersaturations required for intracellular formation of bubbles generally are at least equal to and probably higher than the bubble nucleation thresholds for water or aqueous solutions.

Four microbiological assay procedures (Escherichia coli, Lactobacillus leichmanni, Euglena gracilis, and Ochromonas malhamensis) for the vitamin B(12) content of rat liver and kidney tissue were compared. Significant differences in the growth response for portions of the same sample based on the same standard preparations were noted. The liver values with E. coli were strikingly higher than those obtained with the other microorganisms, about which there was rather general agreement. With kidney tissue, E. coli again gave higher values, but not as significantly high as those obtained with L. leichmannii and E. gracilis. O. malhamensis values were significantly lower. These differences were greater than were expected and could not be explained by the differences of specificity and sensitivity of the assay organisms. Several methods of sample preparation, the possible presence of an alkali-heat-stable factor, different methods of sterilization of the sample plus medium, and different chromatographic procedures were explored in an attempt to find explanations for the variations in growth response, with the thought that these various methods would clarify in what form or forms the free and bound vitamin B(12) might exist in the samples. No clear-cut explanation was found.