In this work, two antifouling polymer brushes were tested at different shear stress conditions to evaluate their performance in reducing the initial adhesion of Escherichia coli. Assays were performed using a parallel plate flow chamber and a shear stress range between 0.005 and 0.056 Pa. These shear stress values are found in different locations in the human body where biomedical devices are placed. The poly(MeOEGMA) and poly(HPMA) brushes were characterized and it was shown that they can reduce initial adhesion up to 90% when compared to glass. Importantly, the performance of these surfaces was not affected by the shear stress, which is an indication that they do not collapse under this shear stress range. The brushes displayed a similar behavior despite the differences in their chemical composition and surface energy. Both surfaces have shown ultra-low adsorption of macromolecules from the medium when tested with relevant biological fluids (urine and serum). This indicates that these surfaces can potentially be used in biomedical devices to reduce initial bacterial colonization and eventually reduce biofilm formation on these devices.