Prevention and control of biofilm formation in food processing environment has remained a challenge for food industry in term of food safety.  This study was designed to investigate the efficacy of hydrogen peroxide pre-treatment combination with the regular procedure using daily cleaning in shrimp plant.  Single and mixed species biofilm of Listeria spp., Salmonella spp. and Pseudomonas spp. were used as the model.  In laboratory single biofilm on stainless steel coupons (SS) were formed under nutrient stress and harvested at 3 days and 7 days to assess the 4 cleaning procedures.  The result showed that single biofilms of Listeria and Salmonella were completely eliminated by using 2% alkaline detergent 10 min following with 2 type of QUAT based sanitizers.  However this procedure could eliminate Pseudomonas, high potential biofilm formation, by 3-4 log reduction but 5 log reduction was obtained when replaced with acid detergent.  Then mixed species biofilm study was done on 3 materials, stainless steel, Teflon and rubber, the condition was simulated as continuously 7 days process conditions.  H₂O₂ concentration of 1 and 2% at 5 and 10 min as pre-treatments were subjected to mixed biofilm prior to the regular cleaning procedure.  Hydrogen peroxide 2% as pre-treatment reduced population of bacteria by 6 log (CFU/cm²).  No significant different in pre-treatment with cleaning process between 5 min of 2% H₂O₂ and 10 min of 1% H₂O₂, and mixed biofilm on stainless steel was removed the easiest compare to the others.  Applying hydrogen peroxide as the pre-treatment following with the regular cleaning process in plants needed in removing and controlling biofilm, particular mixed species biofilm.