This study aimed to assess the biofilm formation by Bacillus cereus on two novel surfaces namely: aluminum and cold steel in comparison study with stainless steel and polystyrene. Also, it aimed to study the inhibitory effect of a new strain Pediococcus acidilactici against biofilm formation by B. cereus grown on these surfaces. In this study, B. cereus M50 isolated from milky machine surface was selected as the highest biofilm producer. The number of M50 cells adhered to aluminum and stainless steel surfaces were more than that adhered to polystyrene and cold steel, respectively. The antimicrobial, anti-adhesive and SEM studies revealed that the P. acidilactici P12 culture and its cell free filtrate showed a significant potential inhibition of biofilm formation of M50 on all tested surfaces under different conditions. These results demonstrated that P. acidilactici strain are considered a new biotreatment for biofilm destruction of food borne pathogens, food biopreservation and food safety.