How to live at very low substrate concentration.

Availability of carbon/energy sources and temperature are the two environmental factors that severely restrict heterotrophic growth in most ecosystems. DOC concentrations in ground, drinking and surface waters are typically in the range of 0.5-5 mg/L, but most of this is present in a polymeric, inaccessible form for microbes. Concentrations of microbiologically available carbon compounds (so-called assimilable organic carbon, AOC) are usually in the range of 10-100 μg/L, those of individual sugars or amino acids are not higher than a few μg/L. Until recently microbiologists assumed that such nutrient-poor (oligotrophic) environments are "deserts" for life, and that the majority of bacterial cells seen in the microscope are dead, dormant or at least severely starved. Nevertheless, despite the low concentrations of available carbon compounds, bacterial cell numbers recorded in these environments typically are in the range of 10(5)-10(6) per mL. Over the last years, we have learnt that most of these microbes are perfectly alive, metabolizing and ready to grow when given the chance. Hence, microbes have adapted and developed strategies to cope with this situation. Laboratory studies with pure cultures suggest that bacterial cells have developed two strategies to live under such conditions. The first strategy is to perform a "multivorous" way of life by taking up and metabolizing dozens of different carbon substrates simultaneously (i.e., they are NOT specializing on a particular substrate, which they can take up with very high affinity). This "mixed substrate growth" equips the cell with a kinetic advantage and metabolic flexibility. Simultaneous utilization of a multitude of carbon substrates allows fast growth at minute concentrations of individual substrates. The second strategy is to minimize maintenance requirements (unfortunately we still know little about how this is achieved). Recently, flow cytometry has been employed to study microbial growth in very dilute, nutrient-poor environments. The technique allows fast and easy quantification of microbial growth of natural bacterial communities, including "uncultivable" members, under environmental conditions. When combined with strain-specific fluorescent immunoprobes, this technique allows investigation of the growth and competition of pathogens with the indigenous microbial flora. This method is particularly suited for studying questions concerning microbial growth and survival in drinking water systems.