Bacterial Physiological State in Wastewater: Monitoring Maintenance and Production with Leu/TdR Ratio for Less Pollution

In domestic wastewater, bacterial physiology controls cell production (growth, replication) and cell maintenance, determining how energy is allocated between these two processes. The aim here was to develop a method to quantify these cellular processes so that the bacterial physiological state could be manipulated to lower this source of pollution. We simultaneously used the incorporation of radiolabelled thymidine into DNA (a measure of new cell synthesis) and leucine into protein in wastewater to quantitatively distinguish bacterial growth from maintenance processes. We found that DNA and protein syntheses were coupled in wastewater after substrate enrichment (with glucose or acetate)—balanced growth. Once the substrate was depleted, the two processes became uncoupled—unbalanced growth. In this physiological state, the bacteria were synthesising protein, but fewer bacteria were replicating. More energy was allocated to cell maintenance than replication. A mean Leu/TdR ratio of 7.4 was determined for wastewater and was similar to natural aquatic ecosystems. As the bacterial growth rate decreased, the Leu/TdR ratios increased. We show how the simultaneous measurement of [³H]Leu and [³H]TdR quantitatively distinguishes balanced from unbalanced growth. Low [³H]Leu/[³H]TdR ratios indicated bacteria were physiologically stressed, an ideal state for biological wastewater treatment processes (WWTP) as the bacteria divert more energy to maintenance activities instead of growth. Leu/TdR ratios of 70 have been recorded in natural aquatic ecosystems which suggests WWTP have potential to be manipulated to achieve much higher Leu/TdR ratios than we report here. Changes to plant operation to improve operation efficiency include finding the optimum rate of substrate (pollution supply) or alternating aerobic and anaerobic periods to maximise the Leu/TdR ratio to achieve less biomass production for land disposal and more cost-effective operation that generates less pollution in the effluent.