X-ray Imaging of Native Hospital Wastewater Biofilms to Unravel Structure, Mechanics, and Antibiotic Resistance Dissemination

Biofilms in hospital wastewater are hotspots for antibiotic resistance, yet their three-dimensional structure and mechanics remain poorly understood. These properties determine how resilient biofilms are to treatment and how effectively they disseminate resistance, but conventional imaging requires contrast agents and cannot probe dense native matrices. Using an innovative in situ sampling device, we will grow natural multi-species biofilms on substrates of controlled stiffness and study them by meta-transcriptomics, two-photon microscopy, and high-resolution X-ray phase contrast micro-computed tomography. The synchrotron X-ray method uniquely enables non-destructive quantification of biofilm density and matrix organization at micrometer scale, directly linking structure to function. This will open new routes for designing anti-biofilm surface coatings and strategies to reduce resistance spread in healthcare environments.