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Biofilms are defined as sessile communities of bacteria that form on surfaces and are entrapped in a matrix that they themselves produce. Biofilms cause severe problems in many natural (Ferris et al., 1989; Nyholm et al., 2002), clinical (Nicolle, 2005; Rice, 2006), and industrial settings (Brink et al., 1994; McLean et al., 2001; Wood et al., 2006), while being beneficial for waste water treatment and biofuel production (Wang and Chen, 2009). In addition, the bioremediation of crude oil spills involves a biofilm of oil degrading microbes, potentially supplemented by marine flagellates and ciliates (Gertler et al., 2010). Identifying the environmental conditions that prevent or support biofilm formation, as well as understanding the regulatory pathways that signal these conditions, is a pre-requisite to both, the solving of biofilm-associated problems and the use for beneficial purposes. In a previous study by our laboratory (Prüβ et al., 2010), it was determined that nutrition ranked among the more important environmental factors affecting biofilm-associated biomass in Escherichia coli K-12. The key to this study was a high-throughput experiment, where biofilm biomass was determined in a collection of cell surface organelle and global regulator mutants under a variety of combinations of environmental conditions. The cell surface organelles each represented a distinct phase of biofilm formation (Sauer et al., 2002). Flagella are required for reversible attachment (phase I), curli or type I fimbriae are characteristic of irreversible attachment (phase II), and a polymeric capsule forms the matrix that permits the maturation of the biofilm (phase III). Eventually, flagellated bacteria are released from the biofilm (phase IV). Phases III and IV are particularly problematic for the disease progression. Bacteria that are located deep within the mature biofilm are particularly resistant to antibiotics and dispersed bacteria tend to serve as a reservoir that continuously feed the infection. Please, see Figure 1 for the distinction of biofilm phases. The global regulators included in our previous study (Prüβ et al., 2010) are involved in the co-ordinate expression and synthesis of biofilm-associated cell surface organelles. Many of them are components of two-component systems (2CSTS), each consisting of a histidine kinase and a response regulator (for reviews on 2CSTS signaling, please, see Galperin, 2004; Parkinson, 1993; West & Stock, 2001). In response to an environmental stimulus, the sensor kinase uses ATP as a phosphodonor to auto-phosphorylate at a conserved histidine, then

A Combination of Phenotype MicroArrayTM Technology with the ATP Assay Determines the Nutritional Dependence of Escherichia coli Biofilm Biomass