BACKGROUND: Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), a capnophilic facultative anaerobe, is associated with localized aggressive periodontitis and endocarditis. When grown in broth, the cells begin to die rapidly after overnight growth. The cells also often lose viability on plates within a few days. The aim of this study is to identify the cause of the rapid loss of cell viability. METHODS: Cell viabilities, as measured by colony forming units, were determined for cells obtained from isolated colonies and from the dense part of a streak on plates. The effect of pH on cell viability was determined by growing cells in broth at various initial glucose concentrations and with or without added bicarbonate. RESULTS: A. actinomycetemcomitans cells were highly sensitive to even a mildly acidic pH of ≈6. Because the bacteria grew at a glucose concentration that is commonly used in many laboratories, there was a dramatic decrease in cell viability as the pH went <6, which happened long before the culture reached saturation. This was easily avoided by using a lower initial glucose concentration, and under these conditions, the addition of bicarbonate to the growth medium was not necessary. Cells resuspended in buffer without nutrients lost viability much faster at pH 6 than at a higher pH. On plates, the cell viability was much higher in isolated colonies than in the dense area of the streak. CONCLUSIONS: A. actinomycetemcomitans cells rapidly lost viability at even a mildly acidic pH. The problem was easily rectified by growing cells at a low glucose concentration.
BACKGROUND:Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), a capnophilic facultative anaerobe, is associated with localized aggressive periodontitis and endocarditis. When grown in broth, the cells begin to die rapidly after overnight growth. The cells also often lose viability on plates within a few days. The aim of this study is to identify the cause of the rapid loss of cell viability. METHODS: Cell viabilities, as measured by colony forming units, were determined for cells obtained from isolated colonies and from the dense part of a streak on plates. The effect of pH on cell viability was determined by growing cells in broth at various initial glucose concentrations and with or without added bicarbonate. RESULTS:A. actinomycetemcomitans cells were highly sensitive to even a mildly acidic pH of ≈6. Because the bacteria grew at a glucose concentration that is commonly used in many laboratories, there was a dramatic decrease in cell viability as the pH went <6, which happened long before the culture reached saturation. This was easily avoided by using a lower initial glucose concentration, and under these conditions, the addition of bicarbonate to the growth medium was not necessary. Cells resuspended in buffer without nutrients lost viability much faster at pH 6 than at a higher pH. On plates, the cell viability was much higher in isolated colonies than in the dense area of the streak. CONCLUSIONS:A. actinomycetemcomitans cells rapidly lost viability at even a mildly acidic pH. The problem was easily rectified by growing cells at a low glucose concentration.