Effect of neutral electrolyzed water on biofilm formed by meat-related Listeria monocytogenes: Intraspecies variability and influence of the growth surface material

Listeria monocytogenes raises major challenges for the food industry. Due to its capacity to form biofilms, this pathogen can persist in processing environments and contaminate the final products. Neutral electrolyzed water (NEW) may offer a promising and eco-friendly method for controlling L. monocytogenes biofilms, though current in vitro studies on its antibiofilm activity are limited and often focused on reference strains. In this study, we assessed the effect of NEW on biofilms formed by meat-related and reference L. monocytogenes strains on polystyrene and stainless steel. Forty wild-type strains isolated from meat products and processing environments were firstly screened for their biofilm-forming abilities and classified as weak (30 %; 12/40), moderate (55 %; 22/40), and strong (15 %; 6/40) biofilm producers. Twenty-two wild-type and two reference strains were selected for the eradication assays, performed by treating the biofilms with NEW for 9 minutes of total contact time. In silico functional enrichment analysis and the visualization of biofilms by scanning electron microscopy (SEM) were also performed. The NEW treatment resulted in a greater average reduction of viable cells in biofilms formed on polystyrene (4.3 ± 1.0 log10 CFU/cm2) compared to stainless steel (2.9 ± 2.0 log10 CFU/cm2), and a remarkable intraspecies variability was observed. SEM images revealed higher structural damage on biofilms formed on polystyrene. Functional enrichment analysis suggested that clustered regularly interspaced short palindromic repeats (CRISPR)-associated elements could be involved in resistance to the treatments. NEW could be a promising additional tool to mitigate L. monocytogenes biofilms in meat processing environments, although its effect varied with surface material and strain-specific characteristics.