Quantitative evaluation of cryptococcal pathogenesis and antifungal drugs using a silkworm infection model with Cryptococcus neoformans

• Published in: Journal of applied microbiology 2012, Vol.112 (1), p.138-146
• Main Authors: Matsumoto, Y, Miyazaki, S, Fukunaga, D.H, Shimizu, K, Kawamoto, S, Sekimizu, K
• Format: Article
• Language: English
• Subjects: amphotericin B; Animals; Antifungal Agents - pharmacology; antifungal drugs; antifungal properties; Biological and medical sciences; Bombyx - microbiology; Bombyx mori; Cryptococcus neoformans; Cryptococcus neoformans - drug effects; Cryptococcus neoformans - genetics; Cryptococcus neoformans - pathogenicity; Cryptococcus neoformans - physiology; drugs; fluconazole; flucytosine; Fundamental and applied biological sciences. Psychology; fungi; genes; hemolymph; ketoconazole; lethal dose 50; mammals; Microbial Sensitivity Tests - methods; Microbiology; mutants; novel infection model; pathogenesis; pharmacokinetics; quantitative evaluation; rearing; screening; serotypes; silkworms; Temperature; therapeutics; virulence
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/j.1365-2672.2011.05186.x

Aims: To develop an in vivo system that could quantitatively evaluate the therapeutic effects of antifungal drugs using a silkworm infection model with Cryptococcus neoformans. Methods and Results: Silkworms reared at 37°C died after an injection of viable serotype A C. neoformans fungus into the haemolymph. The serotype A C. neoformans, which is known to have higher mammal pathogenicity than the serotype D, was also more virulent against the silkworm. Furthermore, the deletion mutants of genes gpa1, pka1 and cna1, which are genes known to be necessary for the pathogenesis in mammals, showed an increase in the number of fungal cells necessary to kill half of the silkworm population (LD50 value). Antifungal drugs, amphotericin B, flucytosine, fluconazole and ketoconazole, showed therapeutic effects in silkworms infected with C. neoformans. However, amphotericin B was not therapeutically effective when injected into the silkworm intestine, comparable to the fact that amphotericin B is not absorbed by the intestine in mammals. Conclusions: The silkworm–C. neoformans infection model is useful for evaluating the therapeutic effects of antifungal drugs. Significance and Impact of the Study: The silkworm infection model has various advantages for screening antifungal drug candidates. We can also elucidate the cryptococcal pathogenesis and evaluate the in vivo pharmacokinetics and toxicity of each drug.