Bdellovibrio bacteriovorus, strain 6-5-S, multiplied in the presence of washed suspensions of Escherichia coli, Spirillum serpens VHL, and Bacillus megaterium which had been autoclaved for 5 min at 121C. No intracellular life cycle was observed. Bdellovibrio bacteriovorus was also able to multiply in an extract from autoclaved E. coli cells after the cells had been removed by centrifugation. Growth of the parasite was dependent upon the addition of Ca2+ and Mg2+ to the buffer solution. The growth rate and yield of B. bacteriovorus on autoclaved cells were not affected by the initial concentration of the parasite. During multiplication of B. bacteriovorus, amino acids, amino sugars, and reducing sugars were released into the culture fluid.