Interactions between spoilage bacteria in tri-species biofilms developed under simulated meat processing conditions

The formation of biofilms in the food industry is a major issue, as they are a frequent source of contamination of products, which can result in significant economic losses for processors through spoilage of foods or pose serious health concerns for consumers when foodborne pathogens are present. In this study, experiments were carried out using CDC Biofilm Reactors to produce biofilms on two test surfaces (polystyrene and stainless steel coupons) under a regimen for simulated meat processing conditions (SMPC). This entailed a 12 day regimen of daily cycles of filling the reactors with a meat slurry and letting stand for 16 h, followed by draining and refilling with water for an 8 h period in order to mimic a possible scenario of fluctuating periods of nutrient availability and starvation in a meat processing facility. Strains of Pseudomonas fluorescens, Lactobacillus plantarum and Leuconostoc pseudomesenteroides were used for mono and mixed cultures biofilms as they are relevant spoilage bacteria in the meat processing industry. In monoculture, the viable cell densities (CFU/cm²) of the two lactic acid bacteria species tested were higher for biofilms grown on polystyrene as compared to those obtained on stainless steel, whereas viable cell numbers in P. fluorescens monoculture were surface-independent. Synergistic interactions were demonstrated during growth of multi-species biofilms. Results from experiments where one of the 3 strains was inoculated 24 h before introduction of the other two strains showed increased levels of L. plantarum within biofilms grown on both test surfaces. The model developed here serves as a baseline to study the interactions between potential spoilage bacteria during biofilm development.