To simulate food contact surfaces with pits or cracks, stainless steel plates with grooves (depths between 0.2 and 5 mm) were constructed. These plates were artificially contaminated with Listeria monocytogenes in clean conditions, with organic soiling, or after 14 days of biofilm formation after which inactivation of the pathogen by Suma Tab D4 (sodium dichloroisocyanurate, 240 and 300 mg/liter), Suma Bac D10 (quaternary ammonium compound, 740 mg/liter), and bacteriophage suspension (Listex P100) was determined. Both chemical disinfectants performed well in suspension tests and in clean carrier tests according to the European standard with a reduction of more than 5 and 4 log units, respectively, of Listeria cells after 5 min of contact time. However, for the plates with grooves, the reduction could not meet the standard requirement, although a higher reduction of L. monocytogenes was observed in the shallow grooves compared with the deeper grooves. Furthermore, presence of food residues and biofilm reduced the effect of the disinfectants especially in the deep grooves, which was dependent on type of food substrate. Bacteriophages showed the best antimicrobial effect compared with the chemical disinfectants (sodium dichloroisocyanurate and quaternary ammonium compound) in most cases in the shallow grooves, but not in the deep grooves. The chlorine based disinfectants were usually less effective than quaternary ammonium compound. The results clearly demonstrate that surfaces with grooves influenced the antimicrobial effect of the chemical disinfectants and bacteriophages because the pathogen is protected in the deep grooves. The use of bacteriophages to inactivate pathogens on surfaces could be helpful in limited cases; however, use of large quantities in practice may be costly and phage-resistant strains may develop.