Wiping of surfaces contaminated with pathogenic bacteria is a key strategy for combating the transmission of healthcare associated infections. It is essential to understand the extent to which removal of bacteria is modulated by fiber properties, biocidal liquid impregnation and applied hand pressure. The influence of intrinsic and extrinsic factors on the removal efficiencies of pathogenic bacteria was studied. Nonwoven wipes made of either hydrophobic (polypropylene) or hygroscopic (lyocell) fibers were manufactured and dynamic removal efficiency of bacteria studied. The single most important parameter affecting bacterial removal efficiency was impregnation with biocidal liquid (p < 0.05). For inherently hygroscopic 100% regenerated cellulose (lyocell) wipes impregnated with biocidal liquid, removal of E. coli, S. aureus and E. faecalis improved by increasing the fabric surface density and wiping pressure to their maximal values – 150 g.m –2 and 13.80 kN.m –2 , respectively. For inherently hydrophobic 100% polypropylene nonwoven wipes, the same conditions maximized the removal efficiency of S. aureus, but for E. coli and E. faecalis a reduction in the wiping pressure to 4.68 kN.m –2 was required. Best practice involves the use of higher surface density wipes (150 g.m –2 ) containing regenerated cellulose fibers loaded with liquid biocide, and applied with the greatest possible wiping pressure.