Surface topography and chemistry of food contact substances, and microbial nutrition affect pathogen persistence and symbiosis in cocktail Listeria monocytogenes biofilms

The foodborne pathogen Listeria monocytogenes (L. monocytogenes) can form very persistent biofilms on food contact substances, either in monospecies, or in conjunction with background microflora. Although some studies have investigated how bacterial symbiosis facilitates L. monocytogenes survival in complex cocktail biofilms, very little is known about how this core symbiotic relationship is affected by environmental factors. Here, we investigate how L. monocytogenes persistence is affected by symbiosis with Escherichia coli O157:H7, Pseudomonas fluorescens, and Ralstonia insidiosa, and how these relationships are impacted by environmental factors, including food contact surface topography and chemistry, and microbial nutrition. We find that symbiosis can significantly enhance L. monocytogenes persistence in four-species cocktail biofilms (5.71 +/- 0.03 Log CFU/cm2) by up to 1.9 Log CFU/cm2 compared to monospecies biofilms (3.81 +/- 0.08 Log CFU/cm2). We also report that surface topography and chemistry, microbial nutrition, and symbiont species significantly impact the symbiotic relationships involving L. monocytogenes, revealing the adaptability of bacterial symbiosis to changing environmental conditions and the complex nature of cocktail pathogen biofilms. Antagonistic and synergistic interactions involving pathogens in cocktail biofilms, and factors affecting those interactions, are elucidated and can establish a foundation to study symbiosis and its role in mitigating pathogen persistence in food systems, and helping identify unanticipated increased food safety risks of pathogens due to enhanced persistence.