Consecutive Treatments with a Multicomponent Sanitizer Inactivate Biofilms Formed by Escherichia coli O157:H7 and Salmonella enterica and Remove Biofilm Matrix

Abstract

Many foodborne pathogens, including Escherichia coli O157:H7 and Salmonella enterica, can develop biofilms on contact surfaces at meat processing plants. Owing to the high tolerance of the biofilm cells associated with the three-dimensional biofilm structure and the well-expressed bacterial extracellular polymeric substances, it is a real challenge to completely inactivate and remove mature biofilms, as well as further prevent biofilm reoccurrence and pathogen survival. In the present study, we evaluated the effectiveness of consecutive treatments (10 to 120 min per treatment) by repeatedly applying a multicomponent sanitizer, based on a functional mechanism by synergistic combination of hydrogen peroxide and quaternary ammonia compounds, against biofilms formed by E. coli O157:H7 and S. enterica strains. Biofilms on stainless steel surfaces were treated with 2.5, 5, or 10% (recommended working concentration) of the sanitizer applied as a foam or liquid solution. Our results showed that the multicomponent sanitizer significantly (P < 0.05) reduced the amount of viable biofilm cells at all concentrations, as enumerable bacteria were only detected after low-concentration treatments (2.5 or 5%) with short exposure periods (10 or 20 min per treatment). Treatments with high concentrations (5 or 10%) of the sanitizer, multiple consecutive treatments (2 or 3 treatments), and sufficient exposure time (>60 min per treatment) effectively controlled pathogen survival postsanitization. Examination with a scanning electron microscope showed that treatment with the sanitizer at 5% strength significantly dissolved the connecting extracellular polysaccharide matrix and removed the majority of the biofilm matrix. No intact biofilm structure was detected after the 10% sanitizer treatment; instead, scattered individual bacteria with visibly altered cell morphology were observed. The treated bacteria exhibited indented and distorted shapes with shortened cell length and increased surface roughness, indicating severe cell injury and death. Our observations indicated that consecutive treatments with the multicomponent sanitizer was effective in inactivating E. coli O157:H7 and S. enterica biofilms and preventing pathogen reoccurrence.