This study aimed to investigate the formation and effect of a biofilm on copper heat exchangers in full-scale system conditions. A modified Pedersen device with copper coupons was installed in parallel to a heat exchanger system to investigate several physico-chemical parameters, such as bacterial enumeration, carbohydrate content of exopolymeric substances, weight loss of test/control coupons, Cu concentrations, and corrosion products over ten months. Findings of this study showed that planktonic bacterial cells attach to each other and form a mixed-species biofilm on the copper coupon surface even though copper is toxic to a variety of microorganisms. These results also revealed that the mixed-species biofilm has a corrosive effect on copper surfaces used in cooling water systems despite the presence of biocide and the corrosion inhibitor. Additionally, it was demonstrated that a shock-dosed biocide application increased the corrosion rate on copper surface in a real system. Preventing risk of microbiologically influenced corrosion entails appropriate material selection and proper/regular chemical treatment of cooling systems. The current study provides useful insights through the evaluation of corrosion of materials with microbiological techniques.