Numerical spatio-temporal characterization of Listeria monocytogenes biofilms

• The evolution of biofilms formed by three L. monocytogenes strains is characterized.
• Microscopy image analysis, counting methods and motility tests were used.
• Two stainless steel supports, commonly used in food industry, were considered.
• Results reveal interstrain variability in terms of developed biofilm structures.
• Topography of supports and motility influence biofilm formation and structure.

As the structure of biofilms plays a key role in their resistance and persistence, this work presents for the first time the numerical characterization of the temporal evolution of biofilm structures formed by three Listeria monocytogenes strains on two types of stainless-steel supports, AISI 304 SS No. 2B and AISI 316 SS No. 2R.Counting methods, motility tests, fluorescence microscopy and image analysis were combined to study the dynamic evolution of biofilm formation and structure. Image analysis was performed with several well-known parameters as well as a newly defined parameter to quantify spatio-temporal distribution.The results confirm the interstrain variability of L. monocytogenes species regarding biofilm structure and structure evolution. Two types of biofilm were observed: homogeneous or flat and heterogeneous or clustered. Differences in clusters and in attachment and detachment processes were due mainly to the topography and composition of the two surfaces although an effect due to motility was also found.