cf. CA 55, 19130e.In a continuous culture of S. aureofaciens in a synthetic medium using sucrose as the limiting substrate, changes in dilution rate result in significant differences in chlortetracycline and pigment formation.In the steady state, both chlortetracycline and the pigments are synthesized only at dilution rates >0.05 hr.-1 and are not formed at dilution rates <0.05 hr.-1An explanation was sought for these observations.It was found that the introduction of nonpigmented mycelium into a continuous culture never leads to the production of pigments in the medium or to mycelium pigmentation.When pigmented mycelium from a batch culture was used, the pigment was invariably released into the medium unless sucrose was excluded.The results showed that the loss of ability to synthesize pigments and chlortetracycline is inherent in the cells and unaffected by changes in the medium composition during batch cultivation.This loss is a result of long-term cultivation under conditions prevailing during continuous fermentation at low dilution rates; it is impossible to maintain these conditions in batch culture for longer periods of time.The reason for the partial or complete loss of ability of the strain to produce chlortetracycline and pigments (in continuous culture at low dilution rates) is the selection of nonpigmented part of a heterogeneous population with suppression of growth of the pigmented part.When discussing the term steadystate, the concentration of the organism and basic nutrients may remain unchanged while other important variables do change.Therefore, it is important to state the criterion for the definition of the steady-state.