Biofilms are considered a significant reason for the failure of disinfection strategies in industrial water systems due to their resistance to antimicrobial agents. This study is designed to investigate the anti-biofilm activity of hydrogen peroxide (H2O2) at combinations of temperatures and contact times. For this purpose, an in vitro microtiter plate (MTP)-based model system was used for biofilm formation using Escherichia coli (E. coli) strain FL-Tbz isolated from the water system of a pharmaceutical plant. To investigate the anti-biofilm activity of H2O2, it was added at different concentrations (2–7% v/v) to biofilms and incubated at different temperatures (20–60 °C) for 10–40 min to find effective conditions to eradicate biofilms. Maximum biofilms were formed when bacterial suspensions were incubated at 37 °C for 96 h. The rate of biofilm formation using an environmental isolate was higher than that of standard strain. H2O2 at concentrations of ≥6.25% (v/v) at temperatures of ≥40 °C incubated for ≥25 min significantly eradicated the biofilms.

  • The time-saving and fairly cheap MTP-based model system is selected for the evaluation of the anti-biofilm activity of hydrogen peroxide.

  • Different behavior of pharmaceutical water system isolates of E. coli in in vitro biofilm formation can be seen.

  • H2O2 is an effective anti-biofilm agent in optimum concentration, temperature, and exposure time against E. coli isolated from a pharmaceutical water system.

Graphical Abstract

Graphical Abstract
Graphical Abstract
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