A laboratory microcosm has been designed for the cultivation of bacteria on surfaces subjected to an adjustable supply of fluids. Bacteria are grown as a microbial film on halved premolar teeth, mounted back to back. Synthetic saliva is dropped slowly over the teeth throughout experiments. A nutrient supplement is provided at regular intervals. The drops of fluid retained by the teeth can be sampled for metabolic end-products. Alternatively, a miniature glass electrode may be set into one half of a tooth assembly to monitor the pH continuously at the stagnation site between tooth segments. Up to six replicate culture flasks and six electrodes can be accommodated in a single experiment. Satisfactory electrode performance was maintained during 66 h experiments. In initial 48 h experiments, teeth were inoculated with Streptococcus rattus BHT or 'Streptococcus mitior' LPA-1 in pure culture and provided with 1% (w/v) glucose for 1 h every 6 h. Bacteria produced typical responses to glucose feeds leading to the formation of 'Stephan'-like curves of pH-fall. Under these conditions, 'Strep. mitior' was more acidogenic than Strep. rattus and the pattern of acid production was distinct for each organism.
Human teeth in an artificial mouth were inoculated with Streptococcus mutans BHT, Streptococcus mitior LPA-1, or sequentially with both organisms. Incubation was continued for 90 h. Mixed populations were largest when a nutrient supplement containing 5.0% (w/v) sucrose was supplied. Fewer organisms were recovered from experiments with synthetic saliva only, or when a supplement containing 0.05% (w/v) glucose was available. The inoculation sequence determined the total viable count and a larger population resulted when Strep. mutans was the initial colonizer (P less than 0.01). Strep. mutans was always able to become established even when super-infected on to a 24 h plaque of Strep. mitior. The final proportion of Strep mutans was lower when it was the superinfecting organism and the sucrose (P less than 0.01) or glucose (P less than 0.05) nutrient supplement was provided. This work confirms the importance of inoculation sequence and presence of sugars in plaque accumulation and demonstrates the fundamental role of microbial interactions in this process.
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