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SUMMARY : A self-regulating continuous culture apparatus is described, designed for studying growth and enzyme production in bacteria which require complex media; it might also be suitable for the cultivation of suspended tissue cells. The growth vessel is a tilted cardioid-shaped flask spinning about its long axis. Aeration and mixing are achieved in such a manner that no frothing occurs and no 'anti-foam' agents are required. Adherent bacterial colonies do not form on the walls of the growth vessel. Samples are removed rapidly under aerobic conditions. The basal medium can be automatically augmented with special metabolites at constant concentration without contaminating the main medium reservoir. The apparatus is autoclaved in one unit after assembly. Factors governing the design of similar small-scale apparatus are discussed. The fundamental theory of homocontinuous or capacity flow culture has already been established by numerous authors (Monod, 1950; Novick & Szilard, 1950a, b; Spicer, 1955; Herbert, Elsworth & Telling, 1956), and the principles governing the construction of a self-regulating apparatus for main- taining growing bacterial cultures at constant population density are generally agreed. The three basic requirements for aerobic organisms are: a growth vessel in which the culture is thoroughly mixed and aerated; a reservoir to supply fresh medium to the growth vessel at constant rate; a device to keep the volume of culture in the growth vessel constant, by removing culture at the same rate as the addition of fresh medium, However, since various ancillary devices are necessary if the apparatus is to operate accurately without too much attention, and since most operations must be performed at constant temperature under aseptic conditions, the final assembly is rather more complicated than its simple formulation suggests. Each of the practical difficulties which arise can be resolved in various ways, and numerous assemblies for continuous culture have already been described (see the review by Novick, 1955; also De Haan & Winkler, 1955; Herbert et aZ. 1956), but none of them is the perfect all-purpose equipment. When designing a continuous culture method it is therefore necessary to take into account the particular purpose for which it is to be used. An excellent apparatus for mass-production of bacteria may be quite unsuitable for studies of mutation rates or cell metabolism. The pathogenicity of the organism, its nutritional requirements and growth rate also affect the design of the apparatus. The apparatus described here was designed primarily for studying the induced formation of penicillinase in broth cultures of Bacillus cereus at growth

An Apparatus for the Continuous Culture of Bacteria at Constant Population Density