Study on the Effect of Contrast Agent on Biofilms and Their Visualization in Porous Substrate Using X-ray µCT

Investigation of biofilms and visualization using non-destructive imaging techniques like X-ray μCT has recently gained interest. Biofilms are congregations of microorganisms that attach to surfaces and comprise of microbial cells embedded in extracellular polymeric substances (EPS). They are ubiquitous entities that are commonly found in any non-sterile setting and have direct implications on human health. Methods to visualize them in-situ are highly needed to understand their behaviour (attachment and detachment) inside a substrate. Contrast-enhanced X-ray μCT is a 3D imaging technique that is capable of visualising objects that have very low attenuation contrast. The use of contrast agents in X-ray μCT has been an evolving process, however, the possible toxic effect of these chemical compounds against biofilms has not been studied in detail. In this study, we focus on the toxic effect of contrast agents and study the diffusion and drainage of contrast agents in biofilms. We propose using water-soluble potassium bromide (KBr) as a suitable contrast agent for enhancement of the attenuation coefficient of a monoculture of Pseudomonas fluorescens biofilms inside a porous substrate. At the given concentration, KBr proved to be less bactericidal compared to other commonly used contrast agents and at 5% w/v concentration we were able to clearly distinguish between the biofilm and the porous substrate.