Interactions across bacterial species boundaries are usually influenced by environmental stresses, yet little has been evaluated regarding multifactorial stresses on the fate of dual-species biofilm formation in food industry. In this study, the processing conditions of chilled beef were established as a combination of cold and acid stresses (4 °C and pH 5.4), with pH 7.0 or 25 °C serving as the controls, to investigate the interaction of dual-species biofilm between Pseudomonas fluorescens and Listeria monocytogenes. Dual-species biofilms significantly increased biofilm formation at 72 h under the condition of 25°C-pH7.0 and 25°C-pH5.4 (P < 0.05). Compared with mono-species biofilms, the cell numbers of L. monocytogenes in dual-species biofilms were lower at 25 °C (P < 0.05), however, the adherent cells of L. monocytogenes was higher in dual-species biofilms at 4 °C (P < 0.05). Furthermore, the amount of extracellular polysaccharides and proteins secreted by single P. fluorescens biofilms at 4 °C was more than three times than those at 25 °C. The surface-enhanced Raman spectroscopy further profiled the variability of extracellular polymeric substances (EPS) composition. Additionally, RT-qPCR results revealed an upregulation of biofilm-related and genes in co-culture species. It provides valuable insights into the strategies for removing mixed biofilms under diverse stressful conditions in practical food processing.
Keywords: Food safety; Interspecific interaction; L. monocytogenes; Mixed biofilms.
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