A Silkworm Infection Model for Evaluating In Vivo Biofilm Formation by Pathogenic Fungi.

Experimental animal models are necessary for research on infectious diseases. Generally, mammalian animals, such as mice, are used for infection experiments. However, there are ethical issues associated with conducting infection experiments in mammals. This has made it difficult to perform infection experiments with a large number of individuals. The invertebrate silkworm, Bombyx mori, is gaining attention as a model animal for infection experiments, and silkworm infection models with various pathogens have been established. This review provides information on the use of silkworm infection models for fungal infection research and evaluation of in vivo biofilm formation by pathogenic fungi using a novel silkworm experimental system. Various silkworm infection models with pathogenic fungi have been used for the development of antifungal drugs and the identification of fungal virulence-related genes. Furthermore, a catheter-material-inserted silkworm infection model was established to evaluate biofilm formation in vivo. Silkworm infection models have contributed to research on fungal infections.

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.

Candida albicans Promotes the Antimicrobial Tolerance of Escherichia coli in a Cross-Kingdom Dual-Species Biofilm

Cross-kingdom multi-species biofilms consisting of fungi and bacteria are often resistant to antimicrobial treatment, leading to persistent infections. We evaluated whether the presence of Candida albicans affects the antibacterial tolerance of Escherichia coli in dual-species biofilms and explored the underlying mechanism. We found that the survival of E. coli in the presence of antibacterial drugs was higher in dual-species biofilms compared to single-species biofilms. This tolerance-inducing effect was observed in E. coli biofilms that were treated with a C. albicans culture supernatant. To explore the antibacterial tolerance-inducing factor contained in the culture supernatant and identify the tolerance mechanism, a heated supernatant, a supernatant treated with lyticase, DNase, and proteinase K, or a supernatant added to a drug efflux pump inhibitor were used. However, the tolerance-inducing activity was not lost, indicating the existence of some other mechanisms. Ultrafiltration revealed that the material responsible for tolerance-inducing activity was <10 kDa in size. This factor has not yet been identified and needs further studies to understand the mechanisms of action of this small molecule precisely. Nevertheless, we provide experimental evidence that Candida culture supernatant induces E. coli antibacterial tolerance in biofilms. These findings will guide the development of new treatments for dual-species biofilm infections.