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Methodologies for Studying B. subtilis Biofilms as a Model for Characterizing Small Molecule Biofilm Inhibitors

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Quantifying biofilm propagation on chemically modified surfaces.

Michelle C Halsted¹, Amber N Bible², Jennifer L Morrell-Falvey²

¹The Bredesen Center, University of Tennessee, Knoxville, TN, USA.

|October 28, 2022

Summary

Researchers developed new methods to study bacterial biofilm formation, finding that Pantoea sp. YR343 bacteria form a honeycomb biofilm structure on hydrophobic surfaces. This study quantified biofilm propagation and differences in a flagella-mutant strain.

Keywords:

Biofilm Honeycomb Image analysis Imaging Quantification Quantitative Surface

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Area of Science:

Microbiology
Surface Chemistry
Biotechnology

Background:

Bacterial biofilm formation varies significantly between species, complicating laboratory studies.
Controlling surface chemistry is crucial for understanding early biofilm development.

Purpose of the Study:

To develop and validate novel methods for studying early bacterial biofilm propagation.
To investigate the biofilm formation characteristics of *Pantoea* sp. YR343 on surfaces with controlled chemistry.

Main Methods:

Utilized functionalized silanes to precisely control surface chemistry.
Employed a semi-automated image processing algorithm to quantify biofilm morphology and evolution.
Studied wild-type *Pantoea* sp. YR343 and its flagella-deficient (Δ*fliR*) mutant.

Main Results:

*Pantoea* sp. YR343 exhibits minimal attachment to hydrophilic surfaces.
A distinct "honeycomb" biofilm morphology was observed on hydrophobic surfaces.
Biofilm propagation followed a logarithmic growth pattern, quantified by the image processing algorithm.
The flagella-deficient mutant showed reduced surface attachment compared to the wild-type.

Conclusions:

The developed methodology effectively supports the study of bacterial biofilm formation.
Surface hydrophobicity significantly influences *Pantoea* sp. YR343 biofilm structure.
Flagellar motility plays a role in the initial surface attachment of *Pantoea* sp. YR343.