SummaryA pH-stat fermentor is a continuous cultivator in which the feed rate is controlled to maintain a constant pH, i.e., end-product acid concentration. This fermentor has applicatian to the continuous cultivation of lactic acid-producing organisms in milkbased media. The equations describing the operation of this fermentor are developed. It is shown that, where the limiting substrate is the carbon and energy source, the operation of t h e fermentor is essentially equivalent to that of a turbidostat. In contrast to this, where the carbon and energy source is in excess and growth is limited by another substrate, pH-stat fermentation is stable both in regions of substrate excess, where D = P , ) ,~~, comparable with turbidostat operation, and substrate limitation where D < pCLlllaX, comparable with chemostat operation. These conditions are met in milk-based media. An analysis is presented, allowing the prediction of the degree of substrate limitation, cell density, and dilution rate in a pH-stat fermentor from batch-growth kinetics. This was confirmed using experimental data for the growth of S t r r p t o c w r i i s thermophilirs TS2 and Lactobacillus bulgaricus LB I in skim milk. Stable simultaneous growth of two organisms in continuous culture occurs if their growth rates are determined by separate conditions, so that, at steady state, their growth rates are separately made equal to the dilution rate. It is then predicted, and confirmed by experiment, that a mixed culture of S. fhermophiliis TS2 and L . biilguriciis LB I in a pH-stat continuous fermentor in yogurt mix at pH 5.5 would be stable with t h e growth of L. hirlguricirs LBI being substrate unlimited and t h e fermentor operating with D = p,l,ar for L. bulguric~us LBI, and the growth of S. t l w m~p k i l i i s TS2 being substrate limited so that its growth rate is equal to the existing dilution rate. Finally, it is predicted and confirmed by experiment that if the conditions are altered so that the growth of S. thannophilus TS2 is substrate unlimited the stable association is broken down, the fermentor operates with D approaching for S. thermopliilirs TS2, and L . biilgaricus LBI is washed out to the level maintained by wall growth.'