The rank of the fis gene is relatively constant above a specific

The rank of the fis gene is relatively constant above a specific growth rate of approximately 0.2 h-1, and decreases below this growth rate. The difference in gene rank between rpoS and fis increases with

specific growth rate (Figure 3F). This analysis points to the possibility of inferring growth rate from transcriptomic data. For example, in the drip-flow Torin 1 purchase biofilm the difference in rpoS and fis gene rank was -1135 ± 296 (n = 6, ± SD). From Figure 3F, this difference corresponds to a specific growth rate of approximately 0.08 h-1. Taking the results of Figures 3E and 3F together, it appears as if bacteria in the biofilm were growing very slowly. Oxygen availability limits growth in biofilm In this experimental system, two

potentially limiting substrates for bacterial growth were glucose and oxygen. Cell Cycle inhibitor The composition of the medium used ensured excess nitrogen, phosphorous, sulfur, and other elemental requirements. For example, the molar ratio of ammonium to glucose carbon was 2.3, which provided approximately ten-fold excess nitrogen. There is no basis for anticipating that glucose was limiting in any part of the biofilms that were grown in this study. This can best be appreciated by a simple calculation. As derived by Williamson and McCarty [30], the metabolic substrate that will first be depleted in a biofilm can be determined by calculating the dimensionless quantity D eG S G/D eO2 S O2 Y GO2. This ratio is a measure of the relative diffusive fluxes of glucose and oxygen into the biofilm, where D e denotes the buy Erastin effective diffusion coefficient of the respective substrate in the almost biofilm, S denotes the bulk fluid concentration of the respective substrate, and Y GO2 is the stoichiometric coefficient relating the consumption of glucose and oxygen. In the present case, we take the effective diffusion coefficients of oxygen and glucose to be 1.53 × 10-5 cm2 s-1 and 2.69 × 10-6 cm2 s-1, respectively [31]. The yield coefficient has been carefully measured, in biofilms of this bacterium, and is 2.25 g glucose per g oxygen [32]. With the bulk fluid

concentration of glucose at 200 mg l-1 and the bulk fluid concentration of oxygen at 6 mg l-1, the quantity given by the ratio above has a value of 2.6. This value being greater than 1 means that glucose is provided in excess and that oxygen is the limiting substrate. This interpretation is consistent with the strong expression of oprB in biofilm specimens (Figure 3A) and the analysis shown in Figure 4A. Microelectrode measurements provided direct chemical evidence of reduced oxygen availability (Figure 1). Steep oxygen concentration gradients were measured in the vicinity of the biofilm, with parts of the biofilm experiencing oxygen concentrations of 0.2 mg l-1 or less (Figure 1). These measurements are concordant with the transcriptomic analysis of biofilm bacteria that provides direct biological evidence of oxygen limitation (Figure 3B, Table 3).

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