The Influence of Growth Rate and Nutrient Limitation on the Microbial Composition and Biochemical Properties of a Mixed Culture of Oral Bacteria Grown in a Chemostat

A sample of human dental plaque was homogenized in transport fluid and inoculated simultaneously into a glucose-limited and a glucose-excess chemostat maintained at pH 7.0 and a dilution rate (D) of 0.05 h-1. In an attempt to ensure the establishment of slow-growing bacterial populations, two further inoculations of each chemostat with fresh samples of dental plaque took place before a steady-state was attained at this dilution rate. The dilution rate was increased step-wise to D = 0.6 h-1, and then returned directly to D = 0.05 h-1. Contrary to chemostat theory, microbial communities with a high species diversity were maintained under all of the experimental conditions employed, although not all of the bacterial populations present in the inocula established successfully in the chemostat. At each steady-state the bacteriological composition and biochemical properties (fermentation products, enzyme assays and acid production) of the communities of each chemostat was determined. Higher cell yields and a slightly more diverse community were obtained from the glucose-limited chemostat at all dilution rates. A complex mixture of end products of metabolism was obtained from the glucose-limited chemostat, suggesting amino acid catabolism, while lactate was the predominant acid of the glucose-excess culture. In washed-cell experiments, communities from the glucose-excess chemostat produced the lower terminal pH values following a pulse of glucose, with the lowest pH values occurring at the higher dilution rates. A film of micro-organisms, which accumulated around the neck of the chemostat, was sampled at the end of the experiment. The microbial composition of the films from each chemostat differed markedly, and both were different to the community of the bulk fluid of the respective chemostat. Spirochaetes and a population of yeasts were detected in the films from the glucose-limited and glucose-excess chemostats, respectively. No invertase or glucosyltransferase activity, and little glucoamylase-specific glycogen was detected in the communities from either chemostat, although significant endogenous activity, particularly at high dilution rates, was obtained with washed-cells from the glucose-excess chemostat. The results suggest that the chemostat could make a valuable contribution to the study of the ecology of dental plaque.