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After removing the supernatant and adding 1 mL of scintillation cocktail (Ultima-Gold, Quick-Safe A) to each tube, the samples were counted using a TRI-CARB 2100 TR (Packard TRI-CARB 2100 TR) liquid scintillation counter. Leucine incorporation was converted into carbon assimilation by a conservative factor of 3.1 kg C mol?1 with an isotope dilution factor of 2.0 (Simon and Azam, 1989). Dissolved oxygen (DO) and bacterial respiration rates (BR) Oxygen was measured using the Winkler titration procedure (Carpenter, 1965). Briefly, water was immediately c-Met inhibitor fixed with MnSO4 and KI + NaOH and sealed without headspace in 300 mL Winkler bottles (Wheaton? 227497-11). H2SO4 was later added, and samples were titrated with Na2S2O3 using a Metrohm 785 DMP titrino auto-burette and double platinum electrode (end-point titration precision, �� 1 ��mol L?1) similarly to Kress et al. (2014). Respiration rates were determined by the following equation: Respiration=DO(T0?dark)?DO(T48?dark)48h (2) Where DO(T0 dark) is the initial dissolved oxygen concentration and DO (T48 dark) is the dissolved oxygen concentration after 48 h incubation. We assumed that in all bottle incubations bacterial respiration Rho kinase inhibition (BR) accounted for ~90% of the dark respiration of the entire microbial community (see result and discussion for further details). Bacterial carbon demand (BCD) and bacterial growth efficiency (BGE) BCD was defined as the sum of carbon assimilation measured by bacterial production (BP) and carbon oxidation determined through heterotrophic microbial respiration (BR). Oxygen respiration was converted into carbon consumption assuming a respiratory quotient (RQ) of 1 (del Giorgio and Cole, 1998; Anesio et al., 2003; Smith and Prairie, 2004): BCD=BP+BR (3) BGE was calculated as follows: BGE?(%)=BPBP+BR?x100 (4) TEP concentrations and visualization with associated bacteria Water samples (100 mL) were gently (Non-specific serine/threonine protein kinase spectrophotometrically (Thermo GENESYTM). AB dye was calibrated using GX as a purified polysaccharide (Passow and Alldredge, 1995). A factor of 0.74 was used to convert from GX equivalents to carbon (Engel and Passow, 2001). This conversion factor was used as proxy, since TEP chemical composition is likely to change between different marine and fresh water environments. To visualize TEP with bacterial associations, samples (100 mL) were filtered gently (