Silkworm model of biofilm formation: In vivo evaluation of antimicrobial tolerance of a cross-kingdom dual-species (Escherichia coli and Candida albicans) biofilm on catheter material.

Biofilms are formed by microorganisms and their products on the surface of materials such as medical devices. Biofilm formation protects microorganisms from antimicrobial agents. Bacteria and fungi often form dual-species biofilms on the surfaces of medical devices in clinical settings. An experimental system to evaluate in vivo biofilm formation by the pathogenic fungus Candida albicans was established using silkworms inserted with polyurethane fiber (PF), a catheter material. In the present study, we established an in vivo experimental system using silkworms to evaluate the antimicrobial tolerance of Escherichia coli in single- and dual-species biofilms formed on the surface of the PF. The injection of E. coli into the PF-inserted silkworms led to the formation of a biofilm by E. coli on the surface of the PF. E. coli in the biofilm exhibited tolerance to meropenem (MEPM). Furthermore, when E. coli and C. albicans were co-inoculated into the PF-inserted silkworms, a dual-species biofilm formed on the surface of the PF. E. coli in the dual-species biofilm with C. albicans was more tolerant to MEPM than E. coli in the single-species biofilm. These findings suggest the usefulness of an in vivo experimental system using PF-inserted silkworms to investigate the mechanisms of MEPM tolerance in E. coli in single- and dual-species biofilms.