Disinfectant solutions based on the essential oils of Cymbopogon citratus (D.C.) Stapf. and Cymbopogon nardus (L.) Rendle, applied alone or in combination, and a control disinfectant solution were tested in two phases (3 and 240 h) of biofilm formation by Listeria monocytogenes ATCC 19117 on AISI 304 (#4) stainless steel surface. Disinfectant solutions based on essential oils have effectively reduced the number of surface-adhered cells, especially after 60 min of contact. The disinfectant solutions based on a combination of essential oils were capable of reducing 100% (5.64 Log CFU cm−2) the number of surface-adhered cells after 60 min of contact, at 240 h of biofilm formation. Essential oils of C. citratus and C. nardus, alone or in combination, are new alternatives for disinfection of industrial stainless steel surfaces contaminated by L. monocytogenes.